N-heterocyclic inhibitors of tnf-alpha expression

ABSTRACT

N-heterocyclic compounds that block cytokine production via inhibition of p38 kinase are disclosed. In one embodiment, compounds of the present invention are represented by Formula (I): 
     
       
         
         
             
             
         
       
     
     Methods of production, pharmaceutical compositions and methods of treating conditions associated with inappropriate p38 kinase activity or TNF-α expression utilizing compounds of the present invention are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of Ser. No. 11/485,075, filed Jul. 12,2006 which is a continuation of Ser. No. 10/183,179, filed Jun. 26,2002, which claims priority to provisional Application Ser. No.60/301,020, filed Jun. 26, 2001, the disclosures of which areincorporated herein by reference in their entirety.

FIELD OF THE INVENTION

This invention relates to N-heterocyclic compounds that are effective inblocking cytokine production, and in particular the expression ofTNF-alpha (TNF-α), via inhibition of p38 kinase. Compounds of thepresent invention are useful in the treatment of inflammatory diseasessuch as, for example, rheumatoid arthritis.

BACKGROUND OF THE INVENTION

Overproduction of cytokines such as IL-1 and TNF-α is implicated in awide variety of inflammatory diseases, including rheumatoid arthritis(RA), psoriasis, multiple sclerosis, inflammatory bowel disease,endotoxin shock, osteoporosis, Alzheimer's disease and congestive heartfailure, among others [Henry et al., Drugs Fut., 24:1345-1354 (1999);Salituro et al., Curr. Med. Chem., 6:807-823 (1999)]. There isconvincing evidence in human patients that protein antagonists ofcytokines, such as, for example, monoclonal antibody to TNF-α (Enbrel)[Rankin et al., Br. J. Rheumatol., 34:334-342 (1995)], soluble TNF-αreceptor-Fc fusion protein (Etanercept) [Moreland et al., Ann. Intern.Med., 130:478-486 (1999)] and or IL-1 receptor antagonist [Bresnihan etal., Arthritis Rheum., 41:2196-2204 (1998)], can provide effectivetreatment for chronic inflammatory diseases. As none of the currenttreatments for inflammatory diseases provide complete relief ofsymptoms, and as most current treatments are associated with variousdrawbacks such as side effects, improved methods for treatinginflammatory diseases are desirable.

TNF-α is a protein whose synthesis occurs in many cell types in responseto an external stimulus, such as, for example, a mitogen, an infectiousorganism, or trauma. Signaling from the cell surface to the nucleusproceeds via several intracellular mediators including kinases thatcatalyze phosphorylation of proteins downstream in the signalingcascade. Important mediators for the production of TNF-α cytokine arethe mitogen-activated protein (MAP) kinases, and in particular, p38kinase.

p38 Kinases are activated in response to various stress stimuli,including, but not limited to, proinflammatory cytokines, endotoxin,ultraviolet light, and osmotic shock. Activation of p38 requires dualphosphorylation by upstream MAP kinase kinases (MKK3 and MKK6) onthreonine and tyrosine within a Thr-Gly-Tyr motif, characteristic of p38isozymes.

Four iso-forms of p38 have been described. The α and β forms areexpressed in inflammatory cells and are thought to be key mediators ofTNF-α production. Inhibition of the enzymes p38α and β in cells resultsin reduced levels of expression of TNF-α, and such inhibitors areeffective in animal models of inflammatory disease.

Molecular cloning of human p38α identified two isozymes, which are thesplice variant product of a single gene. Three additional gene productshave subsequently been identified, p38β, p38γ, and p38δ. p38 kinasesphosphorylate and activate the transcription factors, ATF-2, MAX, CHOP,and C/ERPb, suggesting a role of p38 kinases in gene regulation. Inaddition, p38 kinases phosphorylate other protein kinases, such as MAPKactivated protein kinase-2/3 (MAPKAP-K2/3, or MK2/3), andMAP-kinase-interacting kinase 1/2 (MNK1/2). Recently, activation of MK2has been shown to be essential for LPS-induced TNF-α expression[Kotlyarov et al., Nature Cell Biol., 1:94-97 (1999)]. Mice lacking MK2exhibit a 90% reduction in the production of TNF-α and are resistant toshock induced by LPS. The reduction in TNF-α amounts is due not todecreased production of the TNF-α mRNA, but rather to diminishedproduction of the TNF-α protein, suggesting that MK2 regulatesbiosynthesis of TNF-α at a post-transcriptional level.

Ample evidence indicates that the p38 pathway serves an important rolein inflammatory process mediated by IL-1 and TNF-α.

Small molecule inhibitors of p38 are expected to have several advantagesover protein inhibitors of TNF-α or IL-1. p38 inhibitors not only blockthe production of TNF-α and IL-1, but also directly interfere with manyof their secondary biological effects. In addition, small moleculeinhibitors are unlikely to induce immune reaction in patients, and arebelieved active following oral administration.

The present invention provides novel compounds that are potent andselective inhibitors of p38α and β, and as such, are also potentinhibitors of TNF-α expression in human cells. Compounds of the presentinvention are useful in the treatment of p38- and TNF-αexpression-mediated inflammatory and other disorders, including, but notlimited to, bone resorption, graft vs. host reaction, atherosclerosis,arthritis, osteoarthritis, rheumatoid arthritis, gout, psoriasis,topical inflammatory disease states, adult respiratory distresssyndrome, asthma, chronic pulmonary inflammatory disease, cardiacreperfusion injury, renal reperfusion injury, thrombus,glomerulonephritis, Chron's disease, ulcerative colitis, inflammatorybowel disease, multiple sclerosis, endotoxin shock, osteoporosis,Alzheimer's disease, congestive heart failure and cachexia.

SUMMARY OF THE INVENTION

The compounds of the present invention are effective as inhibitors ofinappropriate p38 activity, especially iso forms α and β, and in turn,of cytokine production, and in particular, of cellular TNF-alpha (TNF-α)expression. Accordingly, compounds of the invention are useful for theinhibition, prevention and suppression of various pathologies associatedwith such activity, such as, for example, inflammation, asthma,arthritis, atherosclerosis, multiple sclerosis, psoriasis, autoimmunediseases, Alzeheimers disease and congestive heart failure, amongothers.

In one embodiment, the principles of the present invention provide acompound, including isomers, enantiomers, diastereomers, tautomers,pharmaceutically acceptable salts, prodrugs and solvates thereof,represented by Formula (I):

wherein:

one or two of W, Y and X are ═N—;

one of W, Y and X is selected from ═C—CN, ═C—F, ═C—NO₂, ═C—Br, ═C—NH₂,═C—NHC(O)CH₃ and ═C—Cl;

the remaining W, Y or X is ═CH—;

V is —NR⁵—;

Z is halogen or —N(R¹)(R²);

R¹ and R² are the same or different and are selected from hydrogen,alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl,substituted cycloalkyl, heterocyclyl or substituted heterocyclyl;

R⁵ is hydrogen or alkyl;

R⁶ is

R⁷ is hydrogen, alkyl, substituted alkyl, alkoxy, or halogen;

R⁸ is hydrogen, alkyl, alkyloxy or cyano;

R⁹ is —C(O)R¹⁰ or unsubstituted or substituted heterocyclyl;

R¹⁰ is —N(R³¹)(R³²);

R³¹ and R³² are the same or different and are selected from hydrogen,alkyl, substituted alkyl, alkoxy, aryl, substituted aryl, cycloalkyl,substituted cycloalkyl, heterocyclyl or substituted heterocyclyl;

R¹¹ is hydrogen, halogen, O—R³⁵ or —N(R¹²)(R¹³);

R¹² is hydrogen, alkyl, or substituted alkyl;

R¹³ is —(CH₂)_(m)R¹⁴;

—N(R¹²)(R¹³) taken together may form a heterocyclyl or substitutedheterocyclyl;

m is 0, 1, 2 or 3;

R¹⁴ is hydrogen, alkyl, substituted alkyl, —C(O)N(R³¹)(R³²),—N(R³³)C(O)R³⁴, aryl, substituted aryl, cycloalkyl, substitutedcycloalkyl, heterocyclyl, substituted heterocyclyl, heteroaryl,substituted heteroaryl or

R¹⁵ is hydrogen, alkyl or substituted alkyl;

R¹⁶ is hydrogen or alkyl; or

R³³ is hydrogen, alkyl, or substituted alkyl;

R³⁴ is alkyl, substituted alkyl, aryl or substituted aryl;

R³⁵ is hydrogen or -(lower alkyl)-R³⁶;

R³⁶ is N(R³⁷)(R³⁸);

R³⁷ is hydrogen, alkyl, or substituted alkyl;

R³⁸ is -(substituted alkyl)-R¹⁴; and

N(R³⁷)(R³⁸) taken together may form a heterocyclyl or substitutedheterocyclyl.

Preferred compounds of this invention are those of Formula (I) includinga pharmaceutically acceptable salt thereof

wherein:

one or two of W, Y and X are ═N—;

one of W, Y and X is selected from ═C—CN, ═C—F, ═C—NO₂, ═C—Br, ═C—NH₂,═C—NHC(O)CH₃ and ═C—Cl;

the remaining W, Y or X is ═CH—;

V is —NH—;

Z is —N(R¹)(R²);

R¹ and R² are the same or different and are selected from hydrogen,alkyl or substituted alkyl wherein alkyl is of 1 to 8 carbons;

R⁶ is

R⁷ is hydrogen, alkyl of 1 to 4 carbons, alkoxy of 1 to 4 carbons, orhalogen;

R⁸ is hydrogen;

R⁹ is —C(O)R¹⁰ or unsubstituted or substituted heterocyclyl;

R¹⁰ is —NH₂ or unsubstituted or substituted —NH-alkyl, —NH-alkoxy,—NH-heterocyclyl, —NH-phenyl, or —NH—CH₂-phenyl wherein alkyl and alkoxyare of 1 to 6 carbons;

R¹¹ is hydrogen, halogen, O—R³⁵ or —N(R¹²)(R¹³), wherein N(R¹²)(R¹³)taken together may form a monocyclic heterocyclyl or substitutedheterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional nitrogenatoms or wherein

R¹² is hydrogen;

R¹³ is alkyl of 1 to 4 carbons or

R¹⁵ and R¹⁶ are independently selected from hydrogen and methyl;

R³⁵ is hydrogen or -(lower alkyl)-R³⁶;

R³⁶ is N(R³⁷)(R³⁸);

R³⁷ is hydrogen, alkyl, or substituted alkyl;

R³⁸ is -(substituted alkyl)-R¹⁴; and

N(R³⁷)(R³⁸) taken together may form a heterocyclyl or substitutedheterocyclyl.

The principles of the present invention also provide methods ofinhibiting TNF-α expression in a mammal, wherein the methods compriseadministering to the mammal an effective amount of a compoundrepresented by Formula (I), or a prodrug or salt thereof. As usedherein, inhibiting TNF-α expression is intended to include inhibiting,suppressing and preventing conditions associated with inappropriateTNF-α expression, including, but not limited to, inflammation, asthma,arthritis, atherosclerosis, multiple sclerosis, psoriasis, autoimmunediseases, Alzheimer's disease and congestive heart failure.

The principles of the present invention further provide methods oftreating p38 kinase and TNF-α mediated disorders in a mammal, themethods comprising administering to a mammal in need of such treatment,an effective amount of a compound represented by Formula (I), or aprodrug or salt thereof. As used herein, a p38 kinase mediated disordermeans a disorder associated with inappropriate p38 kinase activity; aTNF-α mediated disorder means a disorder associated with inappropriateTNF-α expression. Such disorders include, but are not limited to,inflammation, asthma, arthritis, atherosclerosis, multiple sclerosis,psoriasis, autoimmune diseases, Alzheimer□s disease and congestive heartfailure.

Accordingly, the compounds of the invention, as well as prodrugs orsalts thereof, may be used in the manufacture of a pharmaceuticalcomposition or medicament for the prophylactic or therapeutic treatmentof disease states in mammals. The compounds of the present invention maybe administered as pharmaceutical compositions as a monotherapy, or incombination with, for example, other anti-inflammatory, e.g. a steroidor NSAID (non-steroidal anti-inflammatory drug) and/or immunosuppressiveagents. Such combination therapies can involve the administration of thevarious pharmaceuticals as a single dosage form or as multiple dosageforms administered simultaneously or sequentially.

Any suitable route of administration may be employed for providing apatient with an effective amount of a compound of the present invention.Suitable routes of administration may include, for example, oral,rectal, nasal, buccal, parenteral (such as, intravenous, intrathecal,subcutaneous, intramuscular, intrasternal, intrahepatic, intralesional,intracranial, intra-articular, and intra-synovial), transdermal (suchas, for example, patches), and the like. Due to their ease ofadministration, oral dosage forms, such as, for example, tablets,troches, dispersions, suspensions, solutions, capsules, soft gelatincapsules, and the like, may be preferred. Administration may also be bycontrolled or sustained release means and delivery devices. Methods forthe preparation of such dosage forms are well known in the art.

Pharmaceutical compositions incorporating compounds of the presentinvention may include excipients, a pharmaceutically acceptable carrier,in addition to other therapeutic ingredients. Excipients such asstarches, sugars, microcrystalline cellulose, diluents, lubricants,binders, coloring agents, flavoring agents, granulating agents,disintegrating agents, and the like may be appropriate depending uponthe route of administration. Because of their ease of administration,tablets and capsules represent the most advantageous oral dosage unitforms. If desired, tablets may be coated by standard aqueous ornonaqueous techniques.

The compounds of the present invention may be used in the form ofpharmaceutically acceptable salts derived from inorganic or organicbases, and hydrates thereof. Included among such base salts are ammoniumsalts, alkali metal salts, such as sodium and potassium salts, alkalineearth metal salts, such as calcium and magnesium salts, salts withorganic bases, such as dicyclohexylamine salts, N-methyl-D-glucamine,and salts with amino acids such as arginine, lysine, and so forth.

DETAILED DESCRIPTION OF THE INVENTION

[1] Thus, in a first embodiment, the present invention provides a novelcompound of Formula (I) including isomers, enantiomers, diastereomers,tautomers, pharmaceutically acceptable salts, prodrugs and solvatesthereof, comprising:

wherein:

one or two of W, Y and X are ═N—;

one of W, Y and X is selected from ═C—CN, ═C—F, ═C—NO₂, ═C—Br, ═C—NH₂,═C—NHC(O)CH₃ and ═C—Cl;

the remaining W, Y or X is ═CH—;

V is —NR⁵—;

Z is halogen or —N(R¹)(R²);

R¹ and R² are the same or different and are selected from hydrogen,alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl,substituted cycloalkyl, heterocyclyl or substituted heterocyclyl;

R⁵ is hydrogen or alkyl;

R⁶ is

R⁷ is hydrogen, alkyl, substituted alkyl, alkoxy, or halogen;

R⁸ is hydrogen, alkyl, alkyloxy or cyano;

R⁹ is —C(O)R¹⁰ or unsubstituted or substituted heterocyclyl;

R¹⁰ is —N(R³¹)(R³²);

R³¹ and R³² are the same or different and are selected from hydrogen,alkyl, substituted alkyl, alkoxy, aryl, substituted aryl, cycloalkyl,substituted cycloalkyl, heterocyclyl or substituted heterocyclyl;

R¹¹ is hydrogen, halogen, O—R³⁵ or —N(R¹²)(R¹³);

R¹² is hydrogen, alkyl, or substituted alkyl;

R¹³ is —(CH₂)_(m)R¹⁴;

—N(R¹²)(R¹³) taken together may form a heterocyclyl or substitutedheterocyclyl;

m is 0, 1, 2 or 3;

R¹⁴ is hydrogen, alkyl, substituted alkyl, —C(O)N(R³¹)(R³²),—N(R³³)C(O)R³⁴, aryl, substituted aryl, cycloalkyl, substitutedcycloalkyl, heterocyclyl, substituted heterocyclyl, heteroaryl,substituted heteroaryl or

R¹⁵ is hydrogen, alkyl or substituted alkyl;

R¹⁶ is hydrogen or alkyl; or

R³³ is hydrogen, alkyl, or substituted alkyl;

R³⁴ is alkyl, substituted alkyl, aryl or substituted aryl;

R³⁵ is hydrogen or -(lower alkyl)-R³⁶;

R³⁶ is N(R³⁷)(R³⁸);

R³⁷ is hydrogen, alkyl, or substituted alkyl;

R³⁸ is -(substituted alkyl)-R¹⁴; and

N(R³⁷)(R³⁸) taken together may form a heterocyclyl or substitutedheterocyclyl.

[2] In a preferred embodiment, the present invention provides a compoundof Formula (I) including isomers, enantiomers, diastereomers, tautomers,pharmaceutically acceptable salts, prodrugs and solvates thereof,

wherein:

one or two of W, Y and X are ═N—;

one of W, Y and X is selected from ═C—CN, ═C—F, ═C—NO₂, ═C—Br, ═C—NH₂,═C—NHC(O)CH₃ and ═C—Cl;

the remaining W, Y or X is ═CH—;

V is —NH—;

Z is —N(R¹)(R²);

R¹ and R² are the same or different and are selected from hydrogen,alkyl or substituted alkyl wherein alkyl is of 1 to 8 carbons;

R⁶ is

R⁷ is hydrogen, alkyl of 1 to 4 carbons, alkoxy of 1 to 4 carbons, orhalogen;

R⁸ is hydrogen;

R⁹ is —C(O)R¹⁰ or unsubstituted or substituted heterocyclyl;

R¹⁰ is —NH₂ or unsubstituted or substituted —NH-alkyl, —NH-alkoxy,—NH-heterocyclyl, —NH-phenyl, or —NH—CH₂-phenyl wherein alkyl and alkoxyare of 1 to 6 carbons;

R¹¹ is hydrogen, halogen, O—R³⁵ or —N(R¹²)(R¹³), wherein N(R¹²)(R¹³)taken together may form a monocyclic heterocyclyl or substitutedheterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional nitrogenatoms or wherein

R¹² is hydrogen;

R¹³ is alkyl of 1 to 4 carbons or

R¹⁵ and R¹⁶ are independently selected from hydrogen and methyl;

R³⁵ is hydrogen or -(lower alkyl)-R³⁶;

R³⁶ is N(R³⁷)(R³⁸);

R³⁷ is hydrogen, alkyl, or substituted alkyl;

R³⁸ is -(substituted alkyl)-R¹⁴; and

N(R³⁷)(R³⁸) taken together may form a heterocyclyl or substitutedheterocyclyl.

[3] In a more preferred embodiment, the present invention provides acompound of Formula (I) including isomers, enantiomers, diastereomers,tautomers, pharmaceutically acceptable salts, prodrugs and solvatesthereof,

wherein:

one or two of W, Y and X are ═N—;

one of W, Y and X is selected from ═C—CN, ═C—F, ═C—NO₂, ═C—Br, ═C—NH₂,═C—NHC(O)CH₃ and ═C—Cl;

the remaining W, Y or X is ═CH—;

V is —NH—;

Z is —N(R¹)(R²);

R¹ and R² are the same or different and are selected from hydrogen oralkyl of 1 to 8 carbons;

R⁶ is

R⁷ is hydrogen, methyl, methoxy, Cl, Br, or F;

R⁸ is hydrogen;

R⁹ is —C(O)R¹⁰ or unsubstituted or substituted heterocyclyl;

R¹⁰ is —NH₂, or unsubstituted or substituted —NH-alkyl, —NH-alkoxy,—NH-phenyl, or —NH—CH₂-phenyl wherein alkyl and alkoxy are of 1 to 6carbons; and

R¹¹ is hydrogen, halogen, O—R³⁵ or —N(R¹²)(R¹³), wherein N(R¹²)(R¹³)taken together form a monocyclic heterocyclyl or substitutedheterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional nitrogenatoms.

[4] In another preferred embodiment, the present invention provides acompound of Formula (I) including isomers, enantiomers, diastereomers,tautomers, pharmaceutically acceptable salts, prodrugs and solvatesthereof,

wherein:

one of W, Y and X is ═N—;

one of W, Y and X is selected from ═C—CN, ═C—F, ═C—NO₂, ═C—Br, ═C—NH₂,═C—NHC(O)CH₃ and ═C—Cl;

the remaining W, Y or X is ═CH—;

V is —NH—;

Z is —N(R¹)(R²);

R¹ and R² are the same or different and are selected from hydrogen oralkyl of 1 to 8 carbons;

R⁶ is

R⁷ is hydrogen, methyl, methoxy, Cl, Br, or F;

R⁸ is hydrogen;

R⁹ is —C(O)R¹⁰ or unsubstituted or substituted heterocyclyl;

R¹⁰ is —NH₂, or unsubstituted or substituted —NH-alkyl, —NH-alkoxy,—NH-phenyl, or —NH—CH₂-phenyl wherein alkyl and alkoxy are of 1 to 6carbons;

R¹¹ is hydrogen, halogen, —O—R³⁵ or —N(R¹²)(R¹³), wherein N(R¹²)(R¹³)taken together form a monocyclic heterocyclyl or substitutedheterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional nitrogenatoms; and

R¹⁵ and R¹⁶ are independently selected from hydrogen and methyl.

[5] In another more preferred embodiment, the present invention providesa compound of Formula (I) including isomers, enantiomers, diastereomers,tautomers, pharmaceutically acceptable salts, prodrugs and solvatesthereof,

wherein:

R¹⁰ is —NH₂, unsubstituted or substituted —NH—CH₃, —NH—C₂H₅, —NH—OCH₃,or —NH—OC₂H₅.

[6] In another more preferred embodiment, the present invention providesa compound of Formula (I) including isomers, enantiomers, diastereomers,tautomers, pharmaceutically acceptable salts, prodrugs and solvatesthereof,

wherein:

R⁹ is unsubstituted or substituted triazole, thiazole, oxadiazole orimidazole.

[7] In another more preferred embodiment, the present invention providesa compound of Formula (I) including isomers, enantiomers, diastereomers,tautomers, pharmaceutically acceptable salts, prodrugs and solvatesthereof,

wherein:

R¹¹ is hydrogen, halogen, —O-(substituted alkyl), —NH-(substitutedalkyl) or

[8] In yet another preferred embodiment, the present invention providesa compound of Formula (I) including isomers, enantiomers, diastereomers,tautomers, pharmaceutically acceptable salts, prodrugs and solvatesthereof,

wherein:

R¹¹ is hydrogen, halogen, —O-(substituted alkyl), —NH-(substitutedalkyl) or

[9] In yet another preferred embodiment, the present invention providesa compound of Formula (I) including isomers, enantiomers, diastereomers,tautomers, pharmaceutically acceptable salts, prodrugs and solvatesthereof,

wherein:

two of W, Y and X are ═N—;

the remaining W, Y or X is selected from ═C—CN, ═C—F, ═C—NO₂, ═C—Br,═C—NH₂, ═C—NHC(O)CH₃ and ═C—Cl;

V is —NH—;

Z is —N(R¹)(R²);

R¹ and R² are the same or different and are selected from hydrogen oralkyl of 1 to 8 carbons;

R⁶ is

R⁷ is hydrogen, methyl, methoxy, Cl, Br, or F;

R⁸ is hydrogen;

R⁹ is —C(O)R¹⁰ or unsubstituted or substituted heterocyclyl;

R¹⁰ is —NH₂, or unsubstituted or substituted —NH-alkyl, —NH-alkoxy,—NH-phenyl, or —NH—CH₂-phenyl wherein alkyl and alkoxy are of 1 to 6carbons;

R¹¹ is hydrogen, halogen, —O—R³⁵ or —N(R¹²)(R¹³), wherein N(R¹²)(R¹³)taken together may form a monocyclic heterocyclyl or substitutedheterocyclyl of 5 to 7 atoms containing 1, 2, or 3 additional nitrogenatoms; and

R¹⁵ and R¹⁶ are independently selected from hydrogen and methyl.

[10] In yet another more preferred embodiment, the present inventionprovides a compound of Formula (I) including isomers, enantiomers,diastereomers, tautomers, pharmaceutically acceptable salts, prodrugsand solvates thereof,

wherein:

R¹⁰ is —NH₂, unsubstituted or substituted —NH—CH₃, —NH—C₂H₅, —NH—OCH₃,or —NH—OC₂H₅.

[11] In yet another more preferred embodiment, the present inventionprovides a compound of Formula (I) including isomers, enantiomers,diastereomers, tautomers, pharmaceutically acceptable salts, prodrugsand solvates thereof,

wherein:

R⁹ is unsubstituted or substituted triazole, thiazole, oxadiazole orimidazole.

[12] In yet another more preferred embodiment, the present inventionprovides a compound of Formula (I) including isomers, enantiomers,diastereomers, tautomers, pharmaceutically acceptable salts, prodrugsand solvates thereof,

wherein:

R¹¹ is hydrogen, halogen, —O-(substituted alkyl), —NH-(substitutedalkyl) or

[13] In yet another more preferred embodiment, the present inventionprovides a compound of Formula (I) including isomers, enantiomers,diastereomers, tautomers, pharmaceutically acceptable salts, prodrugsand solvates thereof,

wherein:

R¹¹ is hydrogen, halogen, —O-(substituted alkyl), —NH-(substitutedalkyl) or

[14] In a second preferred embodiment, the present invention provides apharmaceutical composition comprising as an active ingredient, acompound of the invention or a prodrug or salt thereof, and apharmaceutically acceptable carrier. [15] In a preferred embodiment, thepresent invention provides a pharmaceutical composition furthercomprising one or more additional active ingredients. [16] In a morepreferred embodiment, the present invention provides a pharmaceuticalcomposition wherein the additional active ingredient is ananti-inflammatory compound or an immunosuppressive agent. [17] In a morepreferred embodiment, the present invention provides a pharmaceuticalcomposition wherein the additional active ingredient is chosen from asteroid and an NSAID. [18] In a third embodiment, the present inventionprovides a method of inhibiting TNF-α expression in a mammal comprisingadministering to the mammal an effective amount of the pharmaceuticalcomposition of the invention. [19] In a preferred embodiment, thepresent invention provides a method of treating TNF-α mediated disordercomprising administering to a mammal in need of such treatment, aneffective amount of a pharmaceutical composition of the invention. [20]In a more preferred embodiment, the present invention provides a methodof treating TNF-α mediated disorder, wherein the TNF-α mediated disorderis an inflammatory disorder.

[21] In a more preferred embodiment, the present invention provides amethod of treating TNF-α mediated disorder, wherein the TNF-α mediateddisorder is chosen from bone resorption, graft vs. host reaction,atherosclerosis, arthritis, osteoarthritis, rheumatoid arthritis, gout,psoriasis, topical inflammatory disease states, adult respiratorydistress syndrome, asthma, chronic pulmonary inflammatory disease,cardiac reperfusion injury, renal reperfusion injury, thrombus,glomerulonephritis, Chron's disease, ulcerative colitis, inflammatorybowel disease, multiple sclerosis, endotoxin shock, osteoporosis,Alzheimer's disease, congestive heart failure and cachexia.[22] In a more preferred embodiment, the present invention provides amethod of treating TNF-α mediated disorder wherein the pharmaceuticalcomposition of the invention is administered with one or more additionalanti-inflammatory or immunosuppressive agents as a single dose form oras separate dosage forms.[23] In an even more preferred embodiment, the present inventionprovides a method of treating a condition associated with TNF-αexpression in a mammal comprising administering to a mammal in need ofsuch treatment, an effective amount of a pharmaceutical composition ofthe invention.

[24] In an even more preferred embodiment, the present inventionprovides a method of treating a condition associated with TNF-αexpression in a mammal wherein the condition associated with TNF-αexpression is an inflammatory disorder.

[25] In a more preferred embodiment, the present invention provides amethod of treating a condition associated with TNF-α expression, whereinthe condition associated with TNF-α expression is chosen from boneresorption, graft vs. host reaction, atherosclerosis, arthritis,osteoarthritis, rheumatoid arthritis, gout, psoriasis, topicalinflammatory disease states, adult respiratory distress syndrome,asthma, chronic pulmonary inflammatory disease, cardiac reperfusioninjury, renal reperfusion injury, thrombus, glomerulonephritis, Crohn'sdisease, ulcerative colitis, inflammatory bowel disease, multiplesclerosis, endotoxin shock, osteoporosis, Alzheimer's disease,congestive heart failure and cachexia.[26] In a more preferred embodiment, the present invention provides amethod of treating a condition associated with TNF-α expression whereinthe pharmaceutical composition of the invention is administered with oneor more additional anti-inflammatory or immunosuppressive agents as asingle dose form or as separate dosage forms.[27] In another more preferred embodiment, the present inventionprovides a method of treating a condition associated with p38 kinaseactivity in a mammal comprising administering to a mammal in need ofsuch treatment, an effective amount of a pharmaceutical composition ofthe invention.

[28] In another more preferred embodiment, the present inventionprovides a method of treating a condition associated with p38 kinaseactivity in a mammal wherein the condition associated with p38 activityis an inflammatory disorder.

[29] In a more preferred embodiment, the present invention provides amethod of treating a condition associated with p38 kinase activity ischosen from bone resorption, graft vs. host reaction, atherosclerosis,arthritis, osteoarthritis, rheumatoid arthritis, gout, psoriasis,topical inflammatory disease states, adult respiratory distresssyndrome, asthma, chronic pulmonary inflammatory disease, cardiacreperfusion injury, renal reperfusion injury, thrombus,glomerulonephritis, Chron's disease, ulcerative colitis, inflammatorybowel disease, multiple sclerosis, endotoxin shock, osteoporosis,Alzheimer's disease, congestive heart failure and cachexia.[30] In a more preferred embodiment, the present invention provides amethod of treating a condition associated with p38 activity wherein thepharmaceutical composition of the invention is administered with one ormore additional anti-inflammatory or immunosuppressive agents as asingle dose form or as separate dosage forms.[31] In a fourth embodiment, the present invention provides a compoundincluding isomers, enantiomers, diastereomers, tautomers,pharmaceutically acceptable salts, prodrugs and solvates selected from:

Abbreviations & Definitions

The following terms and abbreviations retain the indicated meaningthroughout this disclosure.

ATP=adenosine triphosphate

cDNA=complementary DNA

DCE=dichloroethylene DCM=dichloromethane=methylene chloride=CH₂Cl₂DIC=diisopropylcarbodiimide DIEA=N,N-diisopropylethylamineDMF=N,N-dimethylformamide DMSO=dimethyl sulfoxide DTT=dithiothreitolEDTA=ethylenediaminetetraacetic acid EIA=enzyme immunoassayELISA=enzyme-linked immunosorbent assay Fmoc=9-fluorenylmethoxycarbonylGST=glutathione S-transferase

-   HOBt=1-hydroxybenzotriazole

LPS=lipopolysaccharide MBP=myelin basic proteinMES=2-(N-morpholino)ethanesulfonic acid

mRNA=messenger RNA

PCR=polymerase chain reaction Pr₂NEt=dipropylethylamine

i-Pr₂NEt=diisopropylethylamine

RPMI=Roswell Park Memorial Institute TBS=t-butyldimethylsilylTFA=trifluoroacetic acid THF=tetrahydrofuran

“Alkyl” is intended to include linear or branched hydrocarbon structuresand combinations thereof of 1 to 20 carbons. “Lower alkyl” means alkylgroups of from 1 to about 10, preferably from 1 to about 8, and morepreferably, from 1 to about 6 carbon atoms. Examples of such radicalsinclude methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl,t-butyl, pentyl, iso-amyl, hexyl, octyl and the like.

“Aryl” means an aromatic hydrocarbon radical of 6 to about 16 carbonatoms, preferably of 6 to about 12 carbon atoms, and more preferably of6 to about 10 carbon atoms. Examples of aryl groups are phenyl, which ispreferred, 1-naphthyl and 2-naphthyl.

“Cycloalkyl” refers to saturated hydrocarbon ring structures of from 3to 12 carbon atoms, and preferably from 3 to 6 carbon atoms. Examplesinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl,adamantyl, and the like. “Lower cycloalkyl” refers to cycloalkyl of 3 to6 carbons.

“Heterocyclyl” refers to saturated, partially saturated or unsaturatedmonocyclic structures of from 3 to 8 atoms, preferably 5 or 6 atoms, andbicyclic structures of 9 or 10 atoms containing one or more carbon atomsand from 1 to 4 heteroatoms chosen from O, N, and S. The point ofattachment of the heterocyclyl structure is at an available carbon ornitrogen atom. Examples include: imidazole, pyridine, indole, thiophene,benzopyranone, thiazole, furan, benzimidazole, quinoline, isoquinoline,quinoxaline, pyrimidine, pyrazine, tetrazole, pyrazole, pyrrolyl,pyridinyl, pyrazolyl, triazolyl, pyrimidinyl, pyridazinyl, oxazolyl,thiazolyl, imidazolyl, indolyl, thiophenyl, furanyl, tetrazolyl,2-pyrrolinyl, 3-pyrrolinyl, pyrrolindinyl, 1,3-dioxolanyl, imidazolinyl,imidazolidinyl, pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl,1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-thiadiazolyl,2H-pyranyl, 4H-pyranyl, piperidinyl, 1,4-dithianyl, thiomorpholinyl,pyrazinyl, piperazinyl, 1,3,5-triazinyl, 1,2,5-trithianyl,benzo(b)thiophenyl, benzimidazolyl, quinolinyl, and the like.

“Alkoxy” means a straight, branched or cyclic hydrocarbon configurationand combinations thereof, including from 1 to 20 carbon atoms,preferably from 1 to 8 carbon atoms, more preferably from 1 to about 4carbon atoms, and an oxygen atom at the point of attachment. Suitablealkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,iso-butoxy, s-butoxy, t-butoxy, cyclopropyloxy, cyclohexyloxy, and thelike. “Lower alkoxy” refers to alkoxy groups having from 1 to 4 carbonatoms. Similarly, “alkylthio” refers to such groups having a sulfur atomat the point of attachment.

“Alkenyl” refers to an unsaturated acyclic hydrocarbon radical in somuch as it contains at least one double bond. “Lower alkenyl” refers tosuch radicals containing from about 2 to about 10 carbon atoms,preferably from about 2 to about 8 carbon atoms and more preferably 2 toabout 6 carbon atoms. Examples of suitable alkenyl radicals includepropenyl, buten-1-yl, isobutenyl, penten-1-yl, 2-methylbuten-1-yl,3-methylbuten-1-yl, hexen-1-yl, hepten-1-yl, and octen-1-yl, and thelike.

“Alkynyl” refers to an unsaturated acyclic hydrocarbon radicalcontaining at least one triple bond. Examples include ethynyl, propynyl,and the like.

“Substituted alkyl” means an alkyl wherein one or more hydrogens,preferably one, two, or three hydrogens, attached to an aliphotic carbonare replaced with a substituent such as —N(R³¹)(R³²), alkoxy, alkylthio,halogen, cyano, carboxyl, hydroxyl, —SO₂-alkyl, —CO₂-alkyl, —C(O)-alkyl,nitro, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,heterocyclyl, substituted heterocyclyl, —C(O)—N(R³¹)(R³²), or—NH—C(O)-alkyl. Examples of such substituent groups include methoxy,ethoxy, propoxy, amino, methylamino, dimethylamino, phenyl naphthyl,chlorine, fluorine, and the like.

“Substituted cycloalkyl” means a cycloalkyl wherein one or morehydrogens, preferably one, two or three hydrogens, attached to a ringcarbon are replaced with a substituent such as alkyl, substituted alkyl,—N(R³¹)(R³²), alkoxy, alkylthio, aryl, substituted aryl, halogen, cyano,carboxyl, hydroxyl, nitro, —SO₂-alkyl, —CO₂-alkyl, —C(O)-alkyl,—C(O)—N(R³¹)R³²), or —NH—C(O)-alkyl. Examples of such groups includemethyl, isopropyl, methoxy, ethoxy, propoxy, amino, methylamino,dimethylamino, phenyl, chlorine, fluorine and the like. Also includedwithin this definition are cycloalkyl rings having a fused aryl,preferably phenyl, or cycloalkyl such as

and the like.

“Substituted aryl” means an aryl wherein one or more hydrogens,preferably one, two or three hydrogens, attached to an aromatic carbonare replaced with a substituent such as alkyl, substituted alkyl,—N(R³¹)(R³²), alkoxy, alkylthio, aryl, substituted aryl, halogen, cyano,nitro, carboxyl, hydroxyl, —SO₂-alkyl, —CO₂-alkyl, —C(O)-alkyl,—C(O)—N(R³¹)(R³²), or —NH—C(O)-alkyl. Examples of such substituentsinclude methyl, isopropyl, methoxy, ethoxy, propoxy, amino, methylamino,dimethylamino, phenyl, chlorine, fluorine, —CO₂CH₃, —C(O)—NH₂, and thelike.

“Substituted heterocyclyl” means a heterocyclyl substituted at one ormore available carbon or nitrogen atoms, preferably at one or two carbonand/or nitrogen atoms, with a substituent such as alkyl, substitutedalkyl, —N(R³¹)(R³²), alkoxy, alkylthio, aryl, substituted aryl, halogen,cyano, nitro, oxo, carboxyl, hydroxyl, —SO₂-alkyl, —CO₂-alkyl,—C(O)-alkyl, —C(O)—N(R³¹)(R³²), or —NH—C(O)-alkyl. Examples of suchgroups include methyl isopropyl, methoxy, ethoxy, propoxy, amino,methylamino, dimethylamino, phenyl, chlorine, fluorine and the like.

“Halogen” is intended to include for example, F, Cl, Br and I.

The term “prodrug” refers to a chemical compound that is converted to anactive agent by metabolic processes in vivo. [See, e.g., N. Boder and J.J. Kaminski, Ann. Rep. Med. Chem. 22:303 (1987) and H. Bundgarrd, Adv.Drug Delivery Rev., 3:39 (1989)]. With regard to the present invention,a prodrug of a compound of Formula I is intended to mean any compoundthat is converted to a compound of Formula I by metabolic processes invivo. The use of prodrugs of compounds of Formula I in any of themethods described herein is contemplated and is intended to be withinthe scope of the invention.

Terminology related to “protected,” “protecting” and/or “deprotecting”functionalities is used throughout this application. Such terminology iswell understood by persons of skill in the art and is used in thecontext of processes which involve sequential treatment with a series ofreagents. In this context, a protecting group refers to a group which isused to mask a functionality during a process step in which it wouldotherwise react, but in which reaction is undesirable. The protectinggroup prevents reaction at that step, but may be subsequently removed toexpose the original functionality. The removal or “deprotection” occursafter the completion of the reaction or reactions in which thefunctionality would interfere. Thus, when a sequence of reagents isspecified, as it is in the processes of the invention, the person ofordinary skill can readily envision those groups that would be suitableas “protecting groups” for the functionalities involved.

In the case of the present invention, the typical functionalities thatmust be protected are amines. Suitable groups for that purpose arediscussed in standard textbooks in the field of chemistry, such asProtective Groups in Organic Synthesis by T. W. Greene [John Wiley &Sons, New York, 1991], which is incorporated herein by reference.Particular attention is drawn to the chapter entitled “Protection forthe Amino Group” (pages 309-405). Preferred protecting groups includeBOC and Fmoc. Exemplary methods for protecting and deprotecting withthese groups are found in Greene and Wuts on pages 318 and 327.

Optical Isomers-Diastereomers-Geometric Isomers

Some of the compounds described herein contain one or more asymmetriccenters and may thus give rise to enantiomers, diastereomers, and otherstereoisometric forms which may be defined in terms of absolutestereochemistry as (R)- or (S)-, or as (D)- or (L)- for amino acids. Thepresent invention is meant to include all such possible diastereomers aswell as their racemic and optically pure forms. Optically active (R)-and (S)- or (D)- and (L)-isomers may be prepared using chiral synthonsor chiral reagents, or optically resolved using conventional techniques.When the compounds described herein contain olefinic double bonds orother centers of geometric asymmetry, and unless specified otherwise, itis intended to include both (E)- and (Z)-geometric isomers. Likewise,all tautomeric forms are intended to be included.

Compounds of the invention which incorporate chiral diamines may beresolved into pairs of enantiomers by known techniques. Where pureenantiomers of starting materials are not commercially available, theymay be obtained by classic resolution, which may employ, for example,fractional crystallization of diastereomeric salts. Compounds of theinvention may have more than one chiral center, for example whereinreductive amination of a homochiral intermediate leads to a mixture ofdiastereomers. Racemic intermediates and compounds of the invention mayalso be resolved by chromatographic separation, such as for example,HPLC using a column loaded with a homochiral support, to yield pureisomeric compounds.

The configuration of any carbon-carbon double bond appearing herein isselected for convenience only and is not intended to designate aparticular configuration; thus a carbon-carbon double bond depictedarbitrarily herein as trans may be cis, trans, or a mixture of the twoin any proportion.

In view of the above definitions, other chemical terms used throughoutthis application can be easily understood by those of skill in the art.Terms may be used alone or in any combination thereof. The preferred andmore preferred chain lengths of the radicals apply to all suchcombinations.

Utility

The compounds of the present invention have demonstrated utility asselective inhibitors of inappropriate p38 kinase activity, and inparticular, isoforms p38α and p38β. As such, compounds of the presentinvention have utility in the treatment of conditions associated withinappropriate p38 kinase activity. Such conditions include diseases inwhich cytokine levels are modulated as a consequence of intracellularsignaling via p38, and in particular, diseases that are associated withan overproduction of such cytokines as Il-1, Il-4, IL-8, and inparticular, TNF-α.

As inhibitors of p-38 kinase activity, compounds of the presentinvention are useful in the treatment and prevention of p-38 mediatedconditions including, but not limited to, inflammatory diseases,autoimmune diseases, destructive bone disorders, proliferativedisorders, angiogenic disorders, infectious diseases, neurodegenerativediseases, viral diseases, allergies, myocardial ischemia,reperfusion/ischemia in stroke, heart attacks, organ hypoxia, vascularhyperplasia, cardiac hypertrophy, thrombin-induced platelet aggregation,and conditions associated with prostaglandin endoperoxidase synthase-2.

Inflammatory diseases which may be treated or prevented include, but arenot limited to, acute pancreatitis, chronic pancreatitis, asthma,allergies and adult respiratory distress syndrome.

Autoimmune diseases which may be treated or prevented include, but arenot limited to, glomerulonephritis, rheumatoid arthritis, systemic lupuserythematosis, scleroderma, chronic thyroiditis, Grave□s disease,autoimmune gastritis, diabetes, autoimmune hemolytic anemia, autoimmuneneutropenia, thrombocytopenia, atopic dermatitis, chronic activehepatitis, myasthenia gravis, multiple sclerosis, inflammatory boweldisease, ulcerative colitis, Crohn's disease, psoriasis, or graft vs.host disease.

Destructive bone disorders which may be treated or prevented include,but are not limited to, osteoporosis, osteoarthritis and multiplemyeloma-related bone disorder.

Proliferative diseases which may be treated or prevented include, butare not limited to, acute myelogenous leukemia, chronic myelogenousleukemia, metastatic melanoma, Kaposi's sarcoma, and multiple myeloma.

Infectious diseases which may be treated or prevented include, but arenot limited to, sepsis, septic shock, and Shigellosis.

Neurodegenerative diseases which may be treated or prevented by thecompounds of this invention include, but are not limited to, Alzheimer'sdisease, Parkinson's disease, cerebral ischemias or neurodegenerativedisease caused by traumatic injury.

Angiogenic disorders which may be treated or prevented include solidtumors, ocular neovasculization, infantile haemangiomas.

Viral diseases which may be treated or prevented include, but are notlimited to, acute hepatitis infection (including hepatitis A, hepatitisB and hepatitis C), HIV infection and CMV retinitis.

In addition, p38 inhibitors of this invention also exhibit inhibition ofthe expression of inducible pro-inflammatory proteins such asprostaglandin endoperoxide synthase-2 (PGHS-2), also referred to ascyclooxygenase-2 (COX-2). Accordingly, additional p38 mediatedconditions include edema, analgesia, fever and pain, such asneuromuscular pain, headache, pain caused by cancer, dental pain andarthritis pain.

As a result of their p38 inhibitory activity, compounds of the presentinvention have utility in the treatment and prevention of diseasesassociated with cytokine production. For example, compounds of thepresent invention are useful in the treatment and prevention of:

Il-1 mediated diseases such as, for example, rheumatoid arthritis,osteoarthritis, stroke, endotoxemia and/or toxic shock syndrome,inflammatory reaction induced by endotoxin, inflammatory bowel disease,tuberculosis, atherosclerosis, muscle degeneration, cachexia, psoriaticarthritis, Reiter's syndrome, gout, traumatic arthritis, rubellaarthritis, acute synovitis, diabetes, pancreatic B-cell disease andAlzheimer's disease;

IL-8 mediated diseases or conditions such as, for example, thosecharacterized by massive neutrophil infiltration, such as psoriasis,inflammatory bowel disease, asthma, cardiac and renal reperfusioninjury, adult respiratory distress syndrome, thrombosis andglomerulonephritis; and

TNF-mediated diseases or conditions such as rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis, gouty arthritis and otherarthritic conditions, sepsis, septic shock syndrome, adult respiratorydistress syndrome, cerebral malaria, chronic pulmonary inflammatorydisease, silicosis, pulmonary sarcoisosis, bone resorption disease,reperfusion injury, graft vs. host reaction, allograft rejections, feverand myalgias due to infection, cachexia secondary to infection, AIDS,ARC or malignancy, meloid formation, scar tissue formation, Crohn'sdisease, ulcerative colitis, pyresis, viral infections, such as HIV,CMV, influenza and herpes; and veterinary viral infections, such aslentivirus infections, including, but not limited to equine infectiousanemia virus; or retro virus infections, including felineimmunodeficiency virus, bovine immunodeficiency virus, or canineimmunodeficiency virus.

The compounds of formula I including a pharmaceutically acceptable saltor hydrate thereof may be administered by any suitable route asdescribed previously to treat the above mentioned diseases andconditions. The method of administration will, of course, vary dependingupon the type of disease being treated. The amount of active compoundadministered will also vary according to the method of administrationand the disease being treated. An effective amount will be within thedosage range of about 0.1 to about 100 mg/kg, preferably about 0.2 toabout 50 mg/kg, in a single or multiple doses administered atappropriate intervals throughout the day.

The IC₅₀ values (concentration required to inhibit 50% of specificbinding) of compounds of the present invention for inhibition of p38activity are below 5 μM. Preferred compounds have an IC₅₀ below 1 μM.

Biological Assays Generation of p38 Kinases

cDNAs of human p38α, β and γ isozymes were cloned by PCR. These cDNAswere subcloned in the pGEX expression vector (Pharmacia). GST-p38 fusionprotein was expressed in E. Coli and purified from bacterial pellets byaffinity chromatography using glutathione agarose. p38 fusion proteinwas activated by incubating with constitutively active MKK6. Active p38was separated from MKK6 by affinity chromatography. Constitutivelyactive MKK6 was generated according to Raingeaud et al. [Mol. Cell.Biol., 1247-1255 (1996)].

TNF-α Production by LPS-Stimulated PBMCS

Heparinized human whole blood was obtained from healthy volunteers.Peripheral blood mononuclear cells (PBMCs) were purified from humanwhole blood by Ficoll-Hypaque density gradient centrifugation andresuspended at a concentration of 5×10⁶/ml in assay medium (RPMI mediumcontaining 10% fetal bovine serum). 50 μl of cell suspension wasincubated with 50 μl of test compound (4× concentration in assay mediumcontaining 0.2% DMSO) in 96 well-tissue culture plates for 5 minutes atroom temperature. 100 μl of LPS (200 ng/ml stock) was then added to thecell suspension and the plate was incubated for 6 hours at 37° C.Following incubation, the culture medium was collected and stored at−20° C. TNFα concentration in the medium was quantified using a standardELISA kit (Pharmingen-San Diego, Calif.). Concentrations of TNFα andIC50 values for test compounds (concentration of compound that inhibitedLPS-stimulated TNFα production by 50%) were calculated by linearregression analysis.

LPS-Induced TNF Production in THP-1 Cells

Human monocytic THP-1 cells were maintained in RPMI 1640 mediumsupplemented with 10% fetal bovine serum. Cells (40,000 cells in 80 μl)were added to wells of 96-well flat-bottomed plates. Tested compounds(10 μl) or vehicle (3% DMSO) were added to wells. Subsequently, LPS(Sigma, #L7261; 10 μl/well) was added to the cells for a finalconcentration of 1 μg/mL. Plates were incubated overnight at 37° C. and5% CO₂. Supernatant (50 μl/well) was harvested for an ELISA assay. TNFwas captured by an anti-human TNF antibody (R&D, #MAB610) which waspre-absorbed in high binding EIA plates (Costar, #3590). Captured TNFwas recognized by a biotinylated anti-human TNF polyclonal antibody(R&D, #BAF210). Streptavidin conjugated with peroxidase was added toeach well, and the activity of peroxidase was quantitated by a peroxidesubstrate kit (Pierce, #34062 and #34006).

p38 Assay

The assays were performed in V-bottomed 96-well plates. The final assayvolume was 60 μl prepared from three 20 μl additions of enzyme,substrates (MBP and ATP) and test compounds in assay buffer (50 mM TrispH 7.5, 10 mM MgCl₂, 50 mM NaCl and 1 mM DTT). Bacterially expressed,activated p38 was pre-incubated with test compounds for 10 min. prior toinitiation of reaction with substrates. The reaction was incubated at25° C. for 45 min. and terminated by adding 5 μl of 0.5 M EDTA to eachsample. The reaction mixture was aspirated onto a pre-wet filtermatusing a Skatron Micro96 Cell Harvester (Skatron, Inc.), then wash withPBS. The filtermat was then dried in a microwave oven for 1 min.,treated with MeltilLex A scintillation wax (Wallac), and counted on aMicrobeta scintillation counter Model 1450 (Wallac). Inhibition datawere analyzed by nonlinear least-squares regression using Prizm(GraphPad Software). The final concentration of reagents in the assaysare ATP, 1 μM; [γ-³³P]ATP, 3 nM; MBP (Sigma, #M1891), 2 μg/well; p38, 10nM; and DMSO, 0.3%.

Methods of Synthesis

General methods of synthesis for compounds of the present invention areillustrated by the following examples. Compounds of the invention may beprepared by standard techniques known in the art, involving bothsolution and solid phase chemistry. Starting materials are commerciallyavailable or may by readily prepared by one of skill in the art withknown methods, or by methods disclosed herein. Specific embodimentsdescribed are presented by way of illustration only, and the inventionis not limited thereto. Modifications and variations in any givematerial or process step will be readily apparent to one of skill in theart and all are to be included within the scope of the invention.

As illustrated in Scheme 1 and Scheme 2, compounds of Formula I whereinV is —NR⁵—; one or two of W, X and Y are N; and each of Z and R¹¹ areattached to the core pyrimidine or pyridine by —N— or —O—, may beprepared from trihalopyrimidine by sequential reactions with threedifferent amines (1, 2, 3), or two different amines (1, 2) and analcohol, and subsequent introduction of an additional substituent on thepyrimidine core. An alternative method of preparation may start fromdihalocyano-methylsulfanyl-pyrimidine (Scheme 2). Preferably, one of theamines will be an aniline and another will be a diamine suitablyprotected on its distal N. The person of skill will recognize that theamines themselves, the sequence of the three substitutions, as well asthe position of the nitrile may be varied, and are not limited by theparticular example shown in Scheme 1 or Scheme 2.

With respect to Formula I of the invention, Amine 1 corresponds to—N(R⁵)(R⁶); Amine 2 corresponds to -Z; and Amine 3 corresponds to —R¹¹and such designations are used interchangeably in the description below.

Preparation of Amines 3-(4-Methyl-3-nitro-phenyl)-4H-[1,2,4]triazole

Hydrogen chloride was bubbled through a solution of3-nitro-p-tolunitrile (0.49 g, 3 mmol) in 40 mL of ethanol at room tempfor 10 min. The solution was continued stirring at room temp for 60 minand the solvent was then evaporated under vacuum to dryness to give awhite solid.

The intermediate so obtained was dissolved in 20 mL of ethanol,neutralized with sodium ethoxide solution and the resulting precipitatewas removed by filtration. To the filtrate was added at room temp formichydrazide (0.2 g, 3 mmol) and the solution was continued stirring atroom temp for 2 h. After removal of volatiles in vacuo, the residue wasdissolved in 30 mL of m-xylene and refluxed at 150° C. for 16 h. Removalof volatiles in vacuo and purification using flash chromatographyafforded 0.26 g of the final product. (Yield: 43%). MS (m/z) calcd forC₉H₈N₄O₂ (MH+) 205.2, found, 205.1.

Coupling of Substituted Pyridines with Amines2-Chloro-6-[(2,2-dimethyl-propyl)-methyl-amino]-5-fluoro-nicotinonitrile

A solution of 2,6-dichloro-3-cyano-5-fluoropyridine (1.0 g, 5.23 mmol),N-methyl-neopentylamine hydrochloride (830 mg, 6.0 mmol) andtriethylamine (1.6 mL) in acetonitrile (20 mL) was stirred at room tempfor 4 hours. Then volatiles were removed in vacuo and the residue waspartitioned between ethyl acetate and water. The organic layer was dried(sodium sulfate) and concentrated in vacuo to afford the product (1.11g, 83%). C₁₂H₁₅ClFN₃ MS m/e=256 (M+H).

Preparation of Substituted Pyrimidines4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde

To phosphoryl chloride (108 mL) chilled in ice bath was addeddimethylformamide (35 mL). The mixture was allowed to stand at 20 degreefor one hour, then 25 g of 2-methylsulfanyl-pyrimidine-4,6-diol wasadded slowly. After 30 minutes, the reaction mixture was heated to 100°C. for 6 hours. The reaction mixture was poured onto crushed ice and theprecipitate was collected by filtration. The crude product was purifiedwith flash chromatography to afford 11.13 g of4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde (Yield=32%).¹H-NMR (300 MHz, CDCl₃): δ 10.50 (s, 1H), 2.76 (s, 3H).

4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde oxime

4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde (7.34 g, 33.09mmol), hydroxylamine hydrochloride (2.31 g, 33.33 mmol), acetic acid(49.6 mL), and water (3.3 mL) were mixed, and heated to 60° C. for 2hours. The reaction mixture was diluted with water and cooled under icebath. The precipitate was collected and dried (Yield=6.41 g, 82%). MS(m/z): 238 (M+H).

4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbonitrile

4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde oxime (7.20 g,30.38 mmol) was added to neat thionyl chloride (29.63 g, 245 mmol), thenthe mixture was heated to reflux for 4 hours. The reaction mixture waspoured onto ice-water. The precipitate of4,6-dichloro-2-methylsulfanyl-pyrimidine-5-carbonitrile was collectedand dried (Yield=6.15 g, 92%). ¹H-NMR (300 MHz, CDCl₃): δ 2.75 (s, 3H).

3-(6-Chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide

4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbonitrile (2.19 g, 10mmol), 3-amino-4,N-dimethyl-benzamide (1.64 g, 10 mmol), and DIEA (1.40g, 18.8 mmol) were mixed in THF (20 mL). The resulting mixture wasstirred at room temperature for overnight. The solvent was evaporatedand the residue was partitioned between ethyl acetate and water. Theorganic layer was concentrated and the crude product was purified byflash chromatography to obtain3-(6-chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide(2.78 g, 80%). MS (m/z): 348 (M+H).

The following examples illustrate preferred embodiments of the presentinvention and do not limit the scope of the present invention, which isdefined in the claims.

Compounds shown in Tables 1 and 2 have been synthesized according to themethods described herein and have been tested in accordance with theprotocols described below. These compounds are provided by way ofillustration only, and the invention is not intended to be limitedthereto. Exemplary syntheses of some compounds are also provided.

TABLE 1 Ex # m/z R_(t) 1

367 4.67 2

 420,422 3

433 3.09 4

405 2.58 5

499 6

466 4.47 7

452 4.22 8

452 4.16 9

424 3.56 10

438 3.94 11

435 12

486 13

432 14

466 15

449 16

466 17

376 18

475 7.3 19

405 6.2 20

335 4.6 21

445 22

496 4.64 23

383 10.3 24

357 25

399 26

396 27

500 11.0 28

366 29

510 4.81 30

509 4.84 31

509 4.28 32

412 6.0 33

467 34

35

405 5.96 36

480 4.67 37

496 5.06 38

494 4.83 39

468 4.69 40

396 41

479 42

495 43

460 5.51 44

494 4.84 45

495 4.46 46

493 4.20 47

422 48

399 7.5 49

 406,409 5.7 50

353 6.0 51

367 6.4 52

413 7.7 53

367 4.53 54

389 55

403 56

387 6.9 57

373 7.0 58

451 59

451 60

451 61

509 12.5 62

495 12.4

TABLE 2 Example#70

Example#71

Example#72

Example#73

Example#74

Example#75

Example#76

Example#77

Example#78

EXAMPLE 70 Synthesis of3-{3-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-5-fluoro-pyridin-2-ylamino}-N-methoxy-4-methyl-benzamide

A mixture of2-chloro-6-[(2,2-dimethyl-propyl)-methyl-amino]-5-fluoro-nicotinonitrile(120 mg, 0.47 mmol), 3-amino-N-methoxy-4-methyl-benzamide (120 mg, 0.66mmol) and potassium fluoride (30 mg, 0.51 mmol) in DMSO (1 mL) washeated to 150° C. overnight. The reaction mixture was allowed to cooldown to room temp and then partitioned between water and ethyl acetate.The organic layer was dried (sodium sulfate) and concentrated underreduced pressure. The product (7.5 mg, 4%) was obtained afterpurification by silica gel chromatography with 30% EtOAc in hexane aseluent. C₂₁H₂₆FN₅O₂ MS m/e=400 (M+H).

EXAMPLE 71 Synthesis of3-{3-Cyano-6-[(2,2-dimethyl-propyl)-amino]-5-fluoro-pyridin-2-ylamino}-N-methoxy-4-methyl-benzamide

(a) Synthesis of2-Chloro-6-[(2,2-dimethyl-propyl)-amino]-5-fluoro-nicotinonitrile

A mixture of 2,6-dichloro-3-cyano-5-fluoropyridine (1.0 g, 5.23 mmol),neopentylamine (530 mg, 6.0 mmol) and triethylamine (1 mL) inacetonitrile (20 mL) was stirred at room temp for 4 h. After the solventwas removed under reduced pressure, the residue was partitioned betweenethyl acetate and water. The organic layer was dried (sodium sulfate)and concentrated under reduced pressure to afford the product (1.11 g,87%). C₁₁H₁₃ClFN₃ MS m/e=242 (M+H).

(b) Synthesis of3-{3-Cyano-6-[(2,2-dimethyl-propyl)-amino]-5-fluoro-pyridin-2-ylamino}-N-methoxy-4-methyl-benzamide

A mixture of2-chloro-6-[(2,2-dimethyl-propyl)-amino]-5-fluoro-nicotinonitrile (150mg, 0.62 mmol), 3-amino-N-methoxy-4-methyl-benzamide (150 mg, 0.83 mmol)and potassium fluoride (30 mg, 0.51 mmol) in DMSO (1 mL) was heated to150° C. overnight. The reaction mixture was allowed to cool to room tempand then partitioned between water and ethyl acetate. The organic layerwas dried (sodium sulfate) and concentrated under reduced pressure. Theproduct (2.5 mg, 1%) was obtained after purification by silica gelchromatography with 30% EtOAc in hexane as eluent. C₂₀H₂₄FN₅O₂ MSm/e=386 (M+H).

EXAMPLE 73 Synthesis of3-[5-Cyano-6-(2,2-dimethyl-propylamino)-3-fluoro-pyridin-2-ylamino]-N-methoxy-4-methyl-benzamide

(a)3-(3-Cyano-6-chloro-5-fluoro-pyridin-2-ylamino)-N-methoxy-4-methyl-benzamide

A mixture of 2,6-dichloro-5-fluoro-nicotinonitrile (830 mg, 4.34 mmol),3-amino-N-methoxy-4-methyl-benzamide (576 mg, 3.2 mmol) andtriethylamine (0.5 mL) in acetonitrile (10 mL) was heated to 70° C.overnight. Then the solvent was removed under reduced pressure and thereaction mixture was partitioned between water and ethyl acetate. Theorganic layer was dried (sodium sulfate) and concentrated under reducedpressure. The product (220 mg, 21%) was isolated after purification bysilica gel chromatography. C₁₅H₁₂ClFN₄O₂ MS m/e=335 (M+H).

(b) Synthesis of3-[5-Cyano-6-(2,2-dimethyl-propylamino)-3-fluoro-pyridin-2-ylamino]-N-methoxy-4-methyl-benzamide

A mixture of3-(6-chloro-5-cyano-3-fluoro-pyridin-2-ylamino)-N-methoxy-4-methyl-benzamide(52 mg, 0.15 mmol), neopentylamine (0.12 mL) and potassium fluoride (12mg) in DMSO (1 mL) was heated to 150° C. overnight. The product (1.1 mg,1.8%) was isolated after purification by HPLC. C₂₀H₂₄FN₅O₂ MS m/e=386(M+H).

EXAMPLE 72 Synthesis of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-3-fluoro-pyridin-2-ylamino}-N-methoxy-4-methyl-benzamide

(a)3-(3-Cyano-6-chloro-5-fluoro-pyridin-2-ylamino)-N-methoxy-4-methyl-benzamide

A mixture of 2,6-dichloro-5-fluoro-nicotinonitrile (830 mg, 4.34 mmol),3-amino-N-methoxy-4-methyl-benzamide (576 mg, 3.2 mmol) andtriethylamine (0.5 mL) in acetonitrile (10 mL) was heated to 70° C.overnight. Then the solvent was removed under reduced pressure and thereaction mixture was partitioned between water and ethyl acetate. Theorganic layer was dried (sodium sulfate) and concentrated under reducedpressure. The product (220 mg, 21%) was isolated after purification bysilica gel chromatography. C₁₅H₁₂ClFN₄O₂ MS m/e=335 (M+H).

(b) Synthesis of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-3-fluoro-pyridin-2-ylamino}-N-methoxy-4-methyl-benzamide

A mixture of3-(6-chloro-5-cyano-3-fluoro-pyridin-2-ylamino)-N-methoxy-4-methyl-benzamide(55.6 mg, 0.166 mmol), N-methyl-neopentylamine (70 mg, 0.511 mmol),diisopropylethylamine (0.1 mL) and potassium fluoride (12 mg) in DMSO (1mL) was heated to 150° C. overnight. The product (3.9 mg, 5.9%) wasisolated after purification by HPLC. C₂₁H₂₆FN₅O₂ MS m/e=400 (M+H).

EXAMPLE 52 Synthesis of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

This compound was prepared according to procedure for the synthesis of3-(6-Chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide.MS (m/z): 413 (M+H).

EXAMPLE 21 Synthesis of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfonyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

To a solution of3-{5-cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(0.20 g, 0.48 mmol) in acetic acid (8 mL) was added a solution ofpotassium permagnate (87 mg, 0.55 mmol) in water (10 mL). The resultingmixture was heated to 50° C. for 10 minutes. The reaction mixture wasthen diluted with water (20 mL), and the product was extracted withethyl acetate. The product was obtained (199 mg) after drying andremoving the solvent. MS (m/z): 445 (M+H).

EXAMPLE 40 Synthesis of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylamino-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfonyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(44 mg, 0.1 mmol) and methylamine (1 mL, 1M in THF) were mixed inp-dioxane (1 mL) in a sealed tube. The mixture was heated to 80° C. forovernight. The solvent was removed in vacuo, and the product (23 mg) waspurified by the silica gel column chromatography. MS (m/z): 396 (M+H).

EXAMPLE 36 Synthesis of3-[3-Cyano-2-[(2,2-dimethyl-propyl)-methyl-amino]6-(1-methyl-piperidin-4-yloxy)pyridin-4-ylamino]-4,N-dimethyl-benzamide

To a portion of 576 mg of 1-methylpiperidine-4-ol (576 mg; 5 mmol) isadded 616 mg of potassium tert.-butoxide (5.5 mmol) followed by 4.0 mLof DMSO. After stirring this mixture at r.t. for 1 h a portion of 1.0 mLof this mixture is added at r.t. to 19 mg of3-{5-cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfonyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(0.043 mmol) on 0.2 mL of DMSO. The mixture was heated at 60° C. for 3h. At r.t. 5 mL of ethyl acetate is added and the organic layer iswashed with brine (1×4 mL). The organic layer is dried (MgSO₄),volatiles are removed in vacuo and the product is purified via reversedphase prep. HPLC. (Yield: 18.6 mg; 0.026 mmol; 56%). MS (m/z): 480(M+H).

EXAMPLE 41 Synthesis of3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide

3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-4,N-dimethyl-benzamidewas synthesized similar to the synthesis of3-{5-cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylamino-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide.MS (m/z): 479 (M+H).

EXAMPLE 42 Synthesis of3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-N-methoxy-4-methyl-benzamide

(a)3-(6-Chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-N-methoxy-4-methyl-benzamide

4,6-Dichloro-2-methylsulfanyl-pyrimidine-5-carbonitrile (2.19 g, 10mmol), 3-amino-N-methoxy-4-methyl-benzamide (1.80 g, 10 mmol) and DIEA(1.9 mL) in THF (80 mL) were stirred at room temperature for overnight.The solvent was removed in vacuo and the product (3.33 g, 92%) wasobtained after purification by silica gel column chromatography.

(b) Synthesis of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide

3-(6-Chloro-5-cyano-2-methylsulfanyl-pyrimidin-4-ylamino)-N-methoxy-4-methyl-benzamide(3.33 g, 9.2 mmol), N-methyl-neopentylamine hydrochloride (2.05 g, 15mmol) and DIEA (3.87 g, 30 mmol) in THF (10 mL) were heated to 60° C.for overnight. The solvent was removed in vacuo and the product (1.75 g)was obtained after purification by silica gel column chromatography.

(c) Synthesis of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfinyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide

To a solution of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide(0.10 g) in ethanol (5 mL) was added the solution of sodium periodate(0.2 g) in water (1 mL). The resulting solution was heated to 70° C. forovernight. The solvent was evaporated and the residue was partitionedbetween water and ethyl acetate. The organic layer was separated, andconcentrated, and the residue was purified by silica gel columnchromatography to afford the sulfoxide product (90 mg). MS (m/z): 445(M+H).

(d) Synthesis of3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-N-methoxy-4-methyl-benzamide

3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfinyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide(40 mg) and 1-methyl-homopiperazine (0.05 mL) in THF (0.5 mL) wereheated in sealed tube at 75° C. for overnight. After the solvent wasremoved in vacuo, the residue was purified by silica gel columnchromatography to afford the product (6.8 mg). MS (m/z): 495 (M+H).

EXAMPLE 32 Synthesis of3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylamino-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide

3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfinyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide(33 mg) and methylamine (2.5 mL, 2 M in THF) were heated in a sealedtube at 75° C. for overnight. After the solvent was removed in vacuo,the residue was purified by silica gel column chromatography to affordthe product (7.3 mg). MS (m/z): 412 (M+H).

EXAMPLE 33 Synthesis of3-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(pyrrolidin-3-ylamino)-pyrimidin-4-ylamino]-N-methoxy-4-methyl-benzamide

3-{5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfinyl-pyrimidin-4-ylamino}-N-methoxy-4-methyl-benzamide(33 mg) and 1-N-Boc-3-(R)-aminopyrrolidine (30 mg), DIEA (0.2 mL) andp-dioxane (2 mL) were heated in a sealed tube at 75° C. for overnight.After the removal of the solvent in vacuo, the product was purified bysilica gel column chromatography and treated with TFA/DCM (1:1) in orderto remove the Boc-group. The product was then converted to hydrochloridesalt by treating it with hydrochloric acid (1 M in ether) (Yield: 20mg). MS (m/z): 567 (M+H).

EXAMPLE 24 Synthesis of3-{5-Amino-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(0.31 g, 0.8 mmol)) was hydrogenated under 40 psi of hydrogen pressurein the presence of 10% Pd/C for 4 hours in Parr instrument. The catalystwas filtered off through celite and the filtrate was concentrated toafford the product (0.24 g, yield 84%). MS (m/z): 357 (M+H).

EXAMPLE 25 Synthesis of3-{5-(Acetylamino)-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-1-benzamide

3-{5-Amino-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(16 mg), acetyl chloride (3.5 mg), triethylamine (0.02 ml) were stirredin methylene chloride (0.2 mL) at room temperature for overnight. Theproduct (2.8 mg) was purified by preparative thin layer chromatography.MS (m/z): 399 (M+H).

EXAMPLE 14 Synthesis of3-{5-Bromo-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

To a solution of3-{6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(0.10 g) in methylene chloride (2 mL), was added sat. aq. sodiumbicarbonate (0.05 mL) and bromine (0.013 mL). The resulting mixture wasstirred at room temperature for 30 minutes, and ethyl acetate (30 mL)and magnesium sulfate (1 g) was added. After filtration andconcentration, the residue was purified by silica gel columnchromatography to afford the product (61.9 mg). MS (m/z): 466 (M+H).

EXAMPLE 15 Synthesis of3-{5-Bromo-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylamino-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-2-methylamino-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(33 mg), aq. sat. sodium bicarbonate (0.05 mL) and bromine (14 mg) werestirred in methylene chloride (1 mL) at room temperature for 4 hours.The product (14 mg) was purified with preparative thin layerchromatography. MS (m/z): 449 (M+1).

EXAMPLE 11 Synthesis of3-[5-Bromo-6-(2,2-dimethyl-propylamino)-2-methylamino-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide

3-[6-(2,2-Dimethyl-propylamino)-2-methylamino-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide(35 mg), aq. sat. sodium bicarbonate (0.05 mL) and bromine (21 mg) werestirred in methylene chloride (1 mL) at room temperature for overnight.The product (4 mg) was purified with preparative thin layerchromatography. MS (m/z): 435 (M+H).

EXAMPLE 49 Synthesis of3-{5-Bromo-6-[(2,2-dimethyl-propyl)-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

(a) 3-(6-Chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide

To a solution of 4,6-dichloropyrimidine (2.0 g, 13.4 mmol) in p-dioxane(50 mL), 3-amino-4,N-dimethyl-benzamide (3.0 g, 18.3 mmol) and DIEA (2.3mL) were added. The resulting mixture was heated to reflux for 4 days.The solvent was removed in vacuo, the residue was taken in water andethyl acetate, and the ethyl acetate layer was separated andconcentrated. The product was purified by silica gel columnchromatography using ethyl acetate:hexane (1:1) as eluent to afford thepale white solid product (1.6 g, yield 43%). MS (m/z): 277 (M+H).

(b) Synthesis of3-[6-(2,2-Dimethyl-propylamino)-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide

To a solution of 3-(6-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide(0.4 g, 1.44 mmol) in DMSO (3 mL), neopentylamine (0.4 mL, 3.39 mmol)was added. The resulting solution was heated at 110° C. for 4 days. Theproduct was purified by silica gel column chromatography using ethylacetate as eluent to afford the product (0.45 g, yield 99%). MS (m/z):328 (M+H).

(c) Synthesis of3-{5-Bromo-6-[(2,2-dimethyl-propyl)-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

To a solution of3-{6-[(2,2-dimethyl-propyl)-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(0.45 g, 1.37 mmol) in methylene chloride (10 mL), was added sat. aq.sodium bicarbonate (2 mL) and bromine (0.07 mL, 1.37 mmol). Theresulting mixture was stirred at room temperature for 18 hours, and thereaction was then diluted with water (20 mL). The organic layer wasseparated and the aqueous layer was extracted with DCM (10 mL). Thecombined organic layer was dried (sodium sulfate), filtered andconcentrated. The residue was purified by silica gel columnchromatography using 2% methanol in DCM as eluent to afford the product(314 mg, yield 56%). MS (m/z): 406 (M+H).

EXAMPLE 2 Synthesis of3-{5-Bromo-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

(a) 3-(6-Chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide

To a solution of 4,6-dichloropyrimidine (2.0 g, 13.4 mmol) in p-dioxane(50 mL), 3-amino-4,N-dimethyl-benzamide (3.0 g, 18.3 mmol) and DIEA (2.3mL) were added. The resulting mixture was heated to reflux for 4 days.The solvent was removed in vacuo, the residue was taken in water andethyl acetate, and the ethyl acetate layer was separated andconcentrated. The product was purified by silica gel columnchromatography using ethyl acetate:hexane (1:1) as eluent to afford thepale white solid product (1.6 g, yield 43%). MS (m/z): 277 (M+H).

(b) Synthesis of3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

To a solution of 3-(6-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide(0.4 g, 1.44 mmol) in DMSO (3 mL), N-methylneopentylamine hydrochloride(0.4 g, 2.9 mmol) and DIEA (0.5 mL, 2.9 mmol) were added. The resultingsolution was heated at 110° C. for 4 days. The product was purified bysilica gel column chromatography using ethyl acetate as eluent to affordthe product (0.46 g, yield 99%). MS (m/z): 342 (M+H).

(c) Synthesis of3-{5-Bromo-6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

To a solution of3-{6-[(2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(0.46 g, 1.34 mmol) in methylene chloride (10 mL), was added sat. aq.sodium bicarbonate (2 mL) and bromine (0.07 mL, 1.37 mmol). Theresulting mixture was stirred at room temperature for 18 hours, and thereaction was then diluted with water (20 mL). The organic layer wasseparated and the aqueous layer was extracted with DCM (10 mL). Thecombined organic layer was dried (sodium sulfate), filtered andconcentrated. The residue was purified by silica gel columnchromatography using 2% methanol in DCM as eluent to afford the product(250 mg, yield 44%). MS (m/z): 420 (M+H).

EXAMPLE 74 Synthesis of3-(R)-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(pyrrolidin-3-ylamino)-pyrimidin-4-ylamino]-4-N-dimethyl-benzamide

(a)3-(R)-[4-[(2,2-Dimethyl-propyl)-methyl-amino]-6-(2-methyl-5-methylcarbamoyl-phenylamino)-pyrimidin-2-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester

A mixture of3-(R)-[4-fluoro-6-(2-methyl-5-methylcarbamoyl-phenylamino)-pyrimidin-2-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester (0.36 g, 0.81 mmol), N-methyl-neopentylaminehydrochloride (411 mg, 3 mmol) and DIEA (0.4 mL) in 1,4-dioxane (0.5 mL)was stirred at 90° C. overnight. After removing the solvent underreduced pressure the desired product (81 mg) was purified by silica gelchromatography. C₂₈H₄₃N₇O₃ MS m/e=526 (M+H).

(b) Synthesis of3-(R)-[5-Cyano-6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(pyrrolidin-3-ylamino)-pyrimidin-4

To a mixture of3-(R)-[4-[(2,2-dimethyl-propyl)-methyl-amino]-6-(2-methyl-5-methylcarbamoyl-phenylamino)-pyrimidin-2-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester (70 mg, 0.0001 mmol) and sodium cyamide (0.044 g,0.89 mmol) in sat. aq. sodium bicarbonate (1 mL) and methylene chloride(3 mL) at room temp was added bromine (0.045 mL, 0.87 mmol). Theresulting mixture was stirred for 16 h at room temp, then diluted withwater and extracted with methylene chloride (2×15 mL). The combinedorganic layer was dried (sodium sulfate), filtered and concentratedunder reduced pressure. This product was then treated with a mixture oftrifluoroacetic acid and methylene chloride (1:1 v/v, 1 mL). Theresulting solution was stirred at room temp for 2 h, then the solventwas removed under reduced pressure and the product was purified by HPLC.C₂₄H₃₄N₈O MS m/e=451 (M+H).

EXAMPLE 75 Synthesis of3-(R)-[5-Cyano-4-[(2,2-dimethyl-propyl)-methyl-amino]-6-(pyrrolidin-3-ylamino)-pyrimidin-2-ylamino]-4-N-dimethyl-benzamide

To a mixture of3-(R)-[6-[(2,2-dimethyl-propyl)-methyl-amino]-2-(2-methyl-5-methylcarbamoyl-phenylamino)-pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester (128 mg, 0.0002 mmol) and sodium cyamide (0.044 g,0.89 mmol) in sat. aq. sodium bicarbonate (1 mL) and methylene chloride(3 mL) was added bromine (0.045 mL, 0.87 mmol). The resulting mixturewas continued stirring at room temp for 16 h, then diluted with waterand extracted with methylene chloride (2×15 mL). The combined organiclayer was dried (sodium sulfate), filtered and concentrated underreduced pressure. This product was then treated with a mixture oftrifluoroacetic acid and methylene chloride (1:1 v/v, 1 mL). Theresulting solution was stirred at room temp for 2 h. The solvent wasremoved under reduced pressure, and the product was purified by HPLC.C₂₄H₃₄N₈O MS m/e=451 (M+H).

EXAMPLE 62 Synthesis of3-[5-Cyano-4-[(2,2-dimethyl-propyl)-methyl-amino]-6-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-2-ylamino]-N-methoxy-4-methyl-benzamide

To a stirred mixture of3-[4-[(2,2-dimethyl-propyl)-methyl-amino]-6-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-2-ylamino]-N-methoxy-4-methyl-benzamide(140 mg, 0.29 mmol) and sodium cyamide (0.044 g, 0.89 mmol) in sat. aq.sodium bicarbonate (1 mL) and methylene chloride (3 mL) was addedbromine (0.045 mL, 0.87 mmol). The resulting mixture was stirred for 16h at room temp, then diluted with water and extracted with methylenechloride (2×15 mL). The combined organic layer was dried (sodiumsulfate), filtered and concentrated under reduced pressure, and theproduct was purified by HPLC. C₂₆H₃₈N₈O₂ MS m/e=495 (M+H). Deprotectionwas performed as described in Example 75.

EXAMPLE 61 Synthesis of3-[5-Cyano-2-[(2,2-dimethyl-propyl)-methyl-amino]-6-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-N-ethoxy-4-methyl-benzamide

To a stirred mixture of3-[2-[(2,2-dimethyl-propyl)-methyl-amino]-6-(4-methyl-[1,4]diazepan-1-yl)-pyrimidin-4-ylamino]-N-ethoxy-4-methyl-benzamide(50 mg, 0.1 mmol) and sodium cyamide (0.044 g, 0.89 mmol) in sat. aq.sodium bicarbonate (1 mL) and methylene chloride (3 mL) was added atroom temp bromine (0.045 mL, 0.87 mmol). The resulting mixture wascontinued stirring for 16 h at room temp. The reaction mixture wasdiluted with water and extracted with methylene chloride (2×15 mL). Thecombined organic layer was dried (sodium sulfate), filtered andconcentrated under reduced pressure, and the product was purified byHPLC. C₂₇H₄₀N₈O₂ MS m/e=509 (M+H).

EXAMPLE 43 Synthesis of4-[(2,2-Dimethyl-propyl)-methyl-amino]-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-2-(pyrrolidin-3(R)-ylamino)-pyrimidine-5-carbonitrile

To a solution of 2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamine (174mg, 1 mmol) in 4 mL of DMF was added 0.17 mL of diisopropylethylamine (1mmol) and 4,6-dichloro-2-methylsulfanyl-pyrimidine-5-carbonitrile (219mg, 1 mmol). The resulting solution was stirred at room temperatureovernight, then partitioned between ethyl acetate and water. The organiclayer was washed with water, brine and dried over MgSO₄. Removal ofvolatiles in vacuo and purification by flash chromatography gave 78 mgof the product (Yield: 22%). MS (m/z) calcd for C₁₅H₁₂ClN₇S (MH+),358.1, found, 358.3.

To a solution of4-chloro-2-methylsulfanyl-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-pyrimidine-5-carbonitrile(71 mg, 0.2 mmol) in 2 mL of n-BuOH was added 137 mg ofN-(2,2-dimethylpropyl)methyl amine HCl salt (1 mmol) and 0.17 mL ofdiisopropylethylamine (1 mmol) and the resulting solution was stirred at60° C. for 24 h. Removal of volatiles in vacuo and purification by flashchromatography gave 68 mg of the product (Yield: 81%). MS (m/z) calcdfor C₂₁H₂₆N₈S (MH+), 423.2, found, 423.3.

To a solution of4-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-pyrimidine-5-carbonitrile(76 mg, 0.18 mmol) in 3 mL of acetic acid was added 0.5 mL waterfollowed by 50 mg of potassium permanganate (0.36 mmol). This solutionwas stirred at 60° C. for 10 minutes, then diluted with water andextracted with ethyl acetate. The organic layer was washed with water,brine and dried over MgSO₄. Removal of volatiles in vacuo gave 58 mg ofthe crude product which was used in the next step without furtherpurification (Yield: 71%). MS (m/z) calcd for C₂₁H₂₆N₈O₂S (MH+), 455.2,found, 455.3.

A solution of4-[(2,2-dimethyl-propyl)-methyl-amino]-2-methanesulfonyl-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-pyrimidine-5-carbonitrile(10 mg, 0.022 mmol) and 1-tert-butoxycarbony-3(R)-amino-pyrrolidine(0.08 g, 0.4 mmol) in 2 mL of acetonitrile was heated with stirring at80° C. for 18 h. Volatiles were the removed in vacuo and the product waspurified by flash chromatography.

This purified product was then dissolved in 3 mL of a solution of 50%TFA in CH₂Cl₂ (v/v) and stirred at room temp for 30 min. Removal ofvolatiles in vacuo and purification via prep. HPLC gave the product asTFA salt. The purified product was then dissolved in 1N HCl (g) indiethyl ether and evaporated to give 0.8 mg of the final product.(Yield: 8%). MS (m/z) calcd for C₂₄H₃₂N₁₀ (MH+), 461.3, found, 461.4.

EXAMPLE 17 Synthesis of4-[(2,2-Dimethyl-propyl)-methyl-amino]-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-pyrimidine-5-carbonitrile

To a solution of4-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-6-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamino]-pyrimidine-5-carbonitrile(60 mg, 0.142 mmol) in 3 mL of 50% ethanol/water (v/v) was added 0.4 mLof 50% Raney-Ni in water solution. The solution was refluxed under argonfor 18 hours. The solution was evaporated under vacuum. The resultingresidue was purified by flash chromatography to afford 5.6 mg of thefinal product (Yield: 10%). MS (m/z) calcd for C₂₀H₂₄N₈ (MH+), 377.2,found, 377.4.

EXAMPLE 80 Synthesis of3-{5-Bromo-2-[2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

(a) 3-(5-Bromo-2-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide

To a solution of 0.38 mL of 5-Bromo-2,4-dichloror-pyrimidine (70 mg; 3.1mmol) in 5 mL of THF at 0° C. is added dropwise a solution of 0.644 mLof N,N-diisopropylethylamine (478 mg; 3.7 mmol) and of 506 mg of3-Amino-4,N-dimethyl-benzamide ((3.1 mmol) in 2 mL of THF. This solutionis continued stirring at 0° C. for 1 h, then at 25° C. for 30 min. Afterremoval of volatiles in vacuo the product was purified via silica gelchromatography (20% ethyl acetate in hexanes) to yield 304 mg of a whitepowder (0.85 mmol; 25% yield). MS (m/z): 355 (M+H).

(b) Synthesis of3-{5-Bromo-2-[2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

A mixture of 51 mg of3-(5-Bromo-2-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide (0.143mmol), 59 mg of (2,2-dimethylpropyl)-methylamine, hydrochloride (0.430mmol) and 0.112 mL of N,N-diisopropylethylamine (83 mg; 0.645 mmol) isheated to 120° C. for 18 h, then allowed to cool to r.t. 5 ml of ethylacetate are added and the organic layer is washed with brine (1×5 ml).The aqueous layer is extracted with ethyl acetate (3×5 ml) and thecombined organic layers are dried (MgSO₄). After removal of volatiles invacuo the product was purified via silica gel chromatography (20% ethylacetate in hexanes) to yield 41 mg of an colorless oil (0.097 mmol;yield: 68%). MS (m/z): 420 (M+H).

EXAMPLE 4 Synthesis of3-[5-Bromo-2-(pyrrolidin-3(R)-(ylamino)-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide

A solution of 22 mg of3-(5-bromo-2-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide (0.06mmol) and 46 mg of 3-amino-pyrrolidine-1-carboxylic acid tert.-butylester (0.247 mmol) is heated at 120° C. in 0.3 mL DMSO for 3 d. Afteraddition of 5 mL of ethyl acetate at r.t. the organic layer is washedwith a sat. solution of NaHCO₃ in water (3×4 mL). The organic layer isdried (MgSO₄), volatiles are removed in vacuo and the product ispurified via prep. HPLC. (Yield: 5.7 mg; 0.013 mmol; 22%). MS (m/z): 444(M⁺); 455 (M+Na).

The purified product is dissolved in 1 mL of MeOH and 3 mL of a 1Nsolution of HCl in diethyl ether is added. The resulting solution isstirred at r.t for 30 min, then volatiles are removed in vacuo and theproduct is purified via prep. HPLC. (Yield: 4.3 mg; 0.0002 mmol; 0.4%).MS (m/z): 405 (M+H).

EXAMPLE 3 Synthesis of3-[Bromo-2-(1-methyl-piperidin-4-yloxy)-pyrimidin-4-ylamino]-4,N-dimethylbenzamide

To a portion of 576 mg of 1-methylpiperidine-4-ol (576 mg; 5 mmol) isadded 616 mg of potassium tert.-butoxide (5.5 mmol) followed by 4.0 mLof DMSO. After stirring this mixture at r.t. for 1 h a portion of 1.0 mLof this mixture is added at r.t. to 21 mg of3-(5-Bromo-2-chloro-pyrimidin-4-ylamino)-4,N-dimethyl-benzamide (0.06mmol). The resulting mixture is heated at 120° C. for 18 h and thenallowed to cool to r.t. A portion of 5 mL of ethyl acetate is added andthe resulting solution is washed with a sat. solution of NaHCO3 in water(3×4 mL). The organic layer is dried (MgSO₄), volatiles are removed invacuo and the product is purified via prep. HPLC. (Yield: 5.7 mg; 0.013mmol; 22%). MS (m/z): 444 (M⁺); 455 (M+Na).

EXAMPLE 53 Synthesis of3-{5-Cyano-2-[2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

A mixture of 15 mg of3-{5-Bromo-2-[2,2-dimethyl-propyl)-methyl-amino]-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(0.036 mmol) and 100 mg of CuCN (1.12 mmol) in 1.0 mL of1-methyl-2-pyrrolidinone is heated to 140° C. for 18 h. The mixture wasallowed to cool to r.t. and 2.0 mL of MeOH were added. After removingthe precipitate by filtration volatiles were evaporated and the productwas purified by prep. HPLC. Yield: 1.7 mg (0.005 mmol; 13%). MS (m/z):367 (M+H).

EXAMPLE 1 Synthesis of3-{5-cyano-4-[-(2,2-dimethyl)(-propyl)-methyl-amino]-pyrimidin-2-ylamino}-4,N-dimethyl-benzamide

To a solution of 1.0 g of 2,4-dichloropyrimidine-5-carbonyl chloride(4.73 mmol) in 5 ml of THF at 0° C. is added dropwise a 0.5 M solutionof NH₃ in 1,4-dioxane. The progress of the amide formation is followedvia HPLC and the solution of NH₃ in 1,4-dioxane is added until all ofthe acid chloride is consumed. Then volatiles are removed in vacuo toyield a white solid.

To the crude product from above is added at r.t. 25 mL of POCl₃ and theresulting mixture is heated to 100° C. for 4 h. Volatiles are removed invacuo, the crude product is absorbed on silica gel and washed off with20% ethyl acetate in hexane to give a white solid.

To 20 mg of the product so obtained in 0.5 mL of THF is added at r.t.0.022 mL of N,N-diisopropylethylamine 16.3 mg; 0.126 mmol) followed by21 mg of 3-amino-4,N-dimethylbenzamide (0.126 mmol). The mixture isstirred at r.t. for 2 h, then 0.5 mL of THF are added followed by 32 mgof (2,2-dimethyl-propyl)-methyl-amine, hydrochloride (0.232 mmol) and0.044 mL of N,N-diisopropylethylamine (32.6 mg; 0.252 mmol). Theresulting mixture is heated at 60° C. for 18 h. Volatiles are removed invacuo and the crude mixture containing two separable regioisomers ispurified via reversed phase prep. HPLC. (Yield: 3.9 mg: 0.011 mmol; 8%).MS (m/z): 367 (M+H).

EXAMPLE 56 Synthesis of3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

(a) (6-Chloro-5-nitro-pyrimidin-4-yl)-(2,2-dimethyl-propyl)-methyl-amine

4,6-Dichloro-5-nitro-pyrimidine (0.20 g, 1.0 mmol),N-methyl-neopentylamine hydrochloride (0.14 g) and DIEA (0.2 mL) werestirred in acetone (5 mL) at 0° C. for 4 hours. The solvent was removedin vacuo and the crude product was used for the next reaction withoutpurification.

(b) Synthesis of3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

(6-Chloro-5-nitro-pyrimidin-4-yl)-(2,2-dimethyl-propyl)-methyl-amine(1.0 mmol) was dissolved in p-dioxane (2 mL), and3-amino-4,N-dimethyl-benzamide (0.2 g, 1.2 mmol) and DIEA (0.3 mL) wereadded. The resulting mixture was heated to 80° C. for overnight. Theproduct (0.31 g, yield 80%) was purified by column silica gelchromatography. MS (m/z): 387 (M+H).

EXAMPLE 26 Synthesis ofN-(2,2-Dimethyl-propyl)-N-methyl-N′-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)-phenyl]-5-nitro-pyrimidine-4,6-diamine

4,6-Dichloro-5-nitro-pyrimidine (193 mg, 1 mmol) was dissolved in 4 mLacetone at 0° C. To the solution was added N-(2,2-dimethylpropyl)methylamine HCl salt (137 mg, 1 mmol) and diisopropylethylamine (0.17 mL, 1mmol). The solution was stirred at 0° C. for 10 minutes, then roomtemperature for 3 hours, and evaporated in vacuo. The crude product waspurified by flash chromatography to afford 180 mg of the product (Yield:69%). MS (m/z) calcd for C₁₀H₁₅ClN₄O₂ (MH+), 259.1, found, 259.3.

A solution of(6-Chloro-5-nitro-pyrimidin-4-yl)-(2,2-dimethyl-propyl)-methyl-amine(185 mg, 0.72 mmol), diisopropylethylamine (0.13 mL, 0.72 mmol) and2-Methyl-5-(4H-[1,2,4]triazol-3-yl)-phenylamine (126 mg, 0.72 mmol) in 3mL of n-BuOH was heated with stirring at 80° C. for 18 h. The solventwas then evaporated in vacuo and the crude product was purified by flashchromatography to afford 87 mg of the product (30%). MS (m/z) calcd forC₁₉H₂₄N₈O₂ (MH+), 397.2, found, 397.3.

EXAMPLE 28 Synthesis ofN-(2,2-Dimethyl-propyl)-N-methyl-N″-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)phenyl]-pyrimidine-4,5,6-triamine

To a solution ofN-(2,2-dimethyl-propyl)-N-methyl-N′-[2-methyl-5-(4H-[1,2,4]triazol-3-yl)phenyl]-5-nitro-pyrimidine-4,6-diamine(20 mg, 0.05 mmol) in 5 mL of methanol was added a catalytic amount of10% Pd/C. The vessel was placed under a hydrogen atmosphere of 20 psifor 1 h at room temperature. The solution was filtered, and the filtratewas evaporated under vacuum to afford 4.6 mg of the product (Yield:25%). MS (m/z) calcd for C₁₉H₂₆N₈ (MH+), 367.2, found, 367.4.

EXAMPLE 12 Synthesis of3-{2-(3-Dimethylamino-propylamino)-6-[(2,2-dimethyl-propyl)-methyl-amino]-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide

2-Methylsulfanyl-pyrimidine-4,6-diol (1.58 g, 10 mmol) was dissolved in10 mL of con. H₂SO₄ at 0° C. To the solution was added 0.84 mL of nitricacid drop-wise. The solution was stirred at 0° C. for 30 minutes, thenroom temperature for 2 h. The solution was poured into ice water. Theyellowish solid precipitated out of the solution was collected, washedwith cold water and dried to afford 400 mg of the product (Yield: 20%).

2-Methylsulfanyl-5-nitro-pyrimidine-4,6-diol (203 mg, 1 mmol) wasdissolved in 4 mL of phosphorus oxychloride. The solution was refluxedat 120° C. for two hours. The solution was evaporated under vacuum. Theoily residue was purified by flash chromatography to afford 80 mg of theproduct (Yield: 35%).

4,6-Dichloro-2-methylsulfanyl-5-nitro-pyrimidine (30 mg, 0.13 mmol) and3-amino-4,N-dimethyl-benzamide (22 mg, 0.13 mmol) were dissolved in 2 mLof THF. The solution was stirred at 0° C. for 30 minutes, and evaporatedunder vacuum. The product thus obtained was directly used for the nextreaction without purification.

The intermediate so obtained was dissolved in 2 mL of THF. To thesolution was added N-(2,2-dimethylpropyl)methyl amine HCl salt (36 mg,0.26 mmol) and diisopropylethylamine (0.05 mL, 0.26 mmol). The solutionwas stirred at room temperature for 1 h, and evaporated under vacuum.The residue was purified by flash chromatography to afford 5.5 mg of theproduct (Yield: 10%, two steps). MS (m/z) calcd for C₂₀H₂₈N₆O₃S (MH+),433.2, found, 433.2.

3-{6-[(2,2-dimethyl-propyl)-methyl-amino]-2-methylsulfanyl-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(24 mg, 0.06 mmol) was dissolved in 1 mL acetic acid. To the solutionwas added two drops of water and potassium permanganate (18 mg, 0.12mmol). The solution was stirred at 60° C. for 10 minutes. The solutionwas diluted with water and extracted with ethyl acetate. The organiclayer was washed with water, brine, dried over MgSO₄, and evaporatedunder vacuum. The residue was further purified by flash chromatographyto afford 4.2 mg of the product (Yield: 15%). MS (m/z) calcd forC₂₀H₂₈N₆O₅S (MH+), 465.2, found, 465.2.

A solution of3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-2-methanesulfonyl-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(4.2 mg, 0.01 mmol) and 3-(Dimethylamino)propylamine (0.2 mL) in 2 mL ofacetonitrile was heated with stirring at 80° C. for 16 h. The solventwas then evaporated under vacuum and the crude product was purified byflash chromatography to afford 2.8 mg of the product (Yield: 64%). MS(m/z) calcd for C₂₄H₃₈N₈O₃ (MH+), 487.3, found, 487.3.

EXAMPLE 5 Synthesis of3-[6-[(2,2-Dimethyl-propyl)-methyl-amino]-2-(1-methyl-piperidin-4-yloxy)-5-nitro-pyrimidin-4-ylamino]-4,N-dimethyl-benzamide

1-Methyl-piperidin-4-ol (56 mg, 0.5 mmol) was dissolved in 2 mL of DMSO.To the solution was added potassium tert-butoxide (56 mg, 0.5 mmol). Thesolution was stirred at room temperature for 1 h. The solution was thenadded to a solution of3-{6-[(2,2-Dimethyl-propyl)-methyl-amino]-2-methanesulfonyl-5-nitro-pyrimidin-4-ylamino}-4,N-dimethyl-benzamide(4 mg, 0.01 mmol) in 0.5 mL of DMSO. The solution was stirred at 80° C.for 3 h, and extracted with ethyl acetate and water. The organic layerwas washed with water, brine, dried over MgSO4, and evaporated undervacuum. The residue was purified by semi-preparative hplc column toafford 0.7 mg of the product (Yield: 16%). MS (m/z) calcd for C₂₅H₃₇N₇O₄(MH+), 500.2, found, 500.1.

Although the present invention has been described in some detail by wayof illustration and example, for purposes of clarity and understanding,it will be apparent that certain changes and modifications may bepracticed within the scope of the appended claims.

1. A compound of Formula (I),

or enantiomers, diastereomers, tautomers, and pharmaceuticallyacceptable salts thereof, wherein: one or two of W, Y and X are ═N—; oneof W, Y and X is selected from ═C—CN, ═C—F, ═C—NO₂, ═C—Br, ═C—NH₂,═C—NHC(O)CH₃ and ═C—Cl; the remaining W, Y or X is ═CH—; V is —NR⁵—; Zis halogen or —N(R¹)(R²); R¹ and R² are the same or different and areselected from hydrogen, alkyl, substituted alkyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl or substitutedheterocyclyl; R⁵ is hydrogen or alkyl; R⁶ is

R⁷ is hydrogen, alkyl, substituted alkyl, alkoxy, or halogen; R⁸ ishydrogen, alkyl, alkyloxy or cyano; R⁹ is —C(O)R¹⁰ or unsubstituted orsubstituted heterocyclyl; R¹⁰ is —N(R³¹)(R³²); R³¹ and R³² are the sameor different and are selected from hydrogen, alkyl, substituted alkyl,alkoxy, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,heterocyclyl or substituted heterocyclyl; R¹¹ is hydrogen, halogen,O—R³⁵ or —N(R¹²)(R¹³); R¹² is hydrogen, alkyl, or substituted alkyl; R¹³is —(CH₂)_(m)R¹⁴; —N(R¹²)(R¹³) taken together may form a heterocyclyl orsubstituted heterocyclyl; m is 0, 1, 2 or 3; R¹⁴ is hydrogen, alkyl,substituted alkyl, —C(O)N(R³¹)(R³²), —N(R³³)C(O)R³⁴, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substitutedheterocyclyl, heteroaryl, substituted heteroaryl or

R¹⁵ is hydrogen, alkyl or substituted alkyl; R¹⁶ is hydrogen or alkyl;or R³³ is hydrogen, alkyl, or substituted alkyl; R³⁴ is alkyl,substituted alkyl, aryl or substituted aryl; R³⁵ is hydrogen or -(loweralkyl)-R³⁶; R³⁶ is N(R³⁷)(R³⁸); R³⁷ is hydrogen, alkyl, or substitutedalkyl; R³⁸ is -(substituted alkyl)-R¹⁴; and N(R³⁷)(R³⁸) taken togethermay form a heterocyclyl or substituted heterocyclyl, provided that whereY and X are each ═N—, then R¹¹ is other than hydrogen or halogen.
 2. Acompound of claim 1 or enantiomers, diastereomers, tautomers, andpharmaceutically acceptable salts thereof, wherein: V is —NH—; Z is—N(R¹)(R²); R¹ and R² are the same or different and are selected fromhydrogen, alkyl, or substituted alkyl wherein alkyl is of 1 to 8carbons; R⁶ is

R⁷ is hydrogen, alkyl of 1 to 4 carbons, alkoxy of 1 to 4 carbons, orhalogen; R⁸ is hydrogen; R⁹ is —C(O)R¹⁰ or unsubstituted or substitutedheterocyclyl; R¹⁰ is —NH₂ or unsubstituted or substituted —NH-alkyl,—NH-alkoxy, —NH-heterocyclyl, —NH-phenyl, or —NH—CH₂-phenyl whereinalkyl and alkoxy are of 1 to 6 carbons; R¹¹ is hydrogen, halogen, O—R³⁵or —N(R¹²)(R¹³), wherein N(R¹²)(R¹³) taken together may form amonocyclic heterocyclyl or substituted heterocyclyl of 5 to 7 atomscontaining 1, 2, or 3 additional nitrogen atoms or wherein R¹² ishydrogen; R¹³ is alkyl of 1 to 4 carbons or

R¹⁵ and R¹⁶ are independently selected from hydrogen and methyl; R³⁵ ishydrogen or -(lower alkyl)-R³⁶; R³⁶ is N(R³⁷)(R³⁸); R³⁷ is hydrogen,alkyl, or substituted alkyl; R³⁸ is -(substituted alkyl)-R¹⁴; andN(R³⁷)(R³⁸) taken together may form a heterocyclyl or substitutedheterocyclyl.
 3. A compound of claim 2 or enantiomers, diastereomers,tautomers, and pharmaceutically acceptable salts thereof, wherein: V is—NH—; Z is —N(R¹)(R²); R¹ and R² are the same or different and areselected from hydrogen or alkyl of 1 to 8 carbons; R⁶ is

R⁷ is hydrogen, methyl, methoxy, Cl, Br, or F; R⁸ is hydrogen; R⁹ is—C(O)R¹⁰ or unsubstituted or substituted heterocyclyl; R¹⁰ is —NH₂, orunsubstituted or substituted —NH-alkyl, —NH-alkoxy, —NH-phenyl, or—NH—CH₂-phenyl wherein alkyl and alkoxy are of 1 to 6 carbons; and R¹¹is hydrogen, halogen, O—R³⁵ or —N(R¹²)(R¹³), wherein N(R¹²)(R¹³) takentogether form a monocyclic heterocyclyl or substituted heterocyclyl of 5to 7 atoms containing 1, 2, or 3 additional nitrogen atoms.
 4. Acompound of claim 3 or enantiomers, diastereomers, tautomers, andpharmaceutically acceptable salts, prodrugs and solvates thereof,wherein: V is —NH—; Z is —N(R¹)(R²); R¹ and R² are the same or differentand are selected from hydrogen or alkyl of 1 to 8 carbons; R⁶ is

R⁷ is hydrogen, methyl, methoxy, Cl, Br, or F; R⁸ is hydrogen; R⁹ is—C(O)R¹⁰ or unsubstituted or substituted heterocyclyl; R¹⁰ is —NH₂, orunsubstituted or substituted —NH-alkyl, —NH-alkoxy, —NH-phenyl, or—NH—CH₂-phenyl wherein alkyl and alkoxy are of 1 to 6 carbons; R¹¹ ishydrogen, halogen, —O—R³⁵ or —N(R¹²)(R¹³), wherein N(R¹²)(R¹³) takentogether form a monocyclic heterocyclyl or substituted heterocyclyl of 5to 7 atoms containing 1, 2, or 3 additional nitrogen atoms; and R¹⁵ andR¹⁶ are independently selected from hydrogen and methyl.
 5. A compoundof claim 4 or enantiomers, diastereomers, tautomers, andpharmaceutically acceptable salts thereof, wherein: R⁹ is —C(O)R¹⁰wherein R¹⁰ is —NH₂, unsubstituted or substituted —NH—CH₃, —NH—C₂H₅,—NH—OCH₃, or —NH—OC₂H₅ or R⁹ is unsubstituted or substituted triazole,thiazole, oxadiazole or imidazole.
 6. (canceled)
 7. A compound of claim5 or enantiomers, diastereomers, tautomers, and pharmaceuticallyacceptable salts thereof, wherein: R¹¹ is hydrogen, halogen,—O-(substituted alkyl), —NH-(substituted alkyl) or


8. (canceled)
 9. (canceled)
 10. (canceled)
 11. (canceled)
 12. (canceled)13. (canceled)
 14. A pharmaceutical composition comprising as an activeingredient, a compound, or a pharmaceutically acceptable salt thereof,according to claim 1, and a pharmaceutically acceptable carrier.
 15. Apharmaceutical composition according to claim 14, further comprising oneor more additional active ingredients which is an anti-inflammatoryagent.
 16. (canceled)
 17. A pharmaceutical composition according toclaim 15, wherein said additional active ingredient is chosen from asteroid and an NSAID.
 18. (canceled)
 19. A method of treating TNF-αmediated disorder, the method comprising administering to a mammal inneed of such treatment, an effective amount of a composition accordingto claim 14 wherein the TNF-α mediated disorder is rheumatoid arthritis,psoriasis, Crohn's disease, osteoarthritis or osteoporosis. 20.(canceled)
 21. (canceled)
 22. The method according to claim 19, whereinsaid composition is administered with one or more additionalanti-inflammatory or immunosuppressive agents as a single dose form oras separate dosage forms.
 23. A method of treating a conditionassociated with TNF-α expression in a mammal, the method comprisingadministering to a mammal in need of such treatment, an effective amountof a composition according to claim
 14. 24. The method according toclaim 23, wherein the condition associated with TNF-α expression is aninflammatory disorder.
 25. The method according to claim 23, wherein thecondition associated with TNF-α expression is chosen from boneresorption, graft vs. host reaction, atherosclerosis, arthritis,osteoarthritis, rheumatoid arthritis, gout, psoriasis, topicalinflammatory disease states, adult respiratory distress syndrome,asthma, chronic pulmonary inflammatory disease, cardiac reperfusioninjury, renal reperfusion injury, thrombus, glomerulonephritis, Crohn'sdisease, ulcerative colitis, inflammatory bowel disease, multiplesclerosis, endotoxin shock, osteoporosis, Alzheimer's disease,congestive heart failure and cachexia.
 26. The method according to claim23 wherein said composition is administered with one or more additionalanti-inflammatory or immunosupressive agents as a single dose form or asseparate dosage forms.
 27. A method of treating a condition associatedwith p38 kinase activity in a mammal, the method comprisingadministering to a mammal in need of such treatment, an effective amountof a composition according to claim
 14. 28. The method according toclaim 27, wherein the condition associated with p38 kinase activity isan inflammatory disorder.
 29. The method according to claim 27, whereinthe condition associated with p38 kinase activity is chosen from boneresorption, graft vs. host reaction, atherosclerosis, arthritis,osteoarthritis, rheumatoid arthritis, gout, psoriasis, topicalinflammatory disease states, adult respiratory distress syndrome,asthma, chronic pulmonary inflammatory disease, cardiac reperfusioninjury, renal reperfusion injury, thrombus, glomerulonephritis, Crohn'sdisease, ulcerative colitis, inflammatory bowel disease, multiplesclerosis, endotoxin shock, osteoporosis, Alzheimer's disease,congestive heart failure and cachexia.
 30. The method according to claim27 wherein said composition is administered with one or more additionalanti-inflammatory or immunosuppressive agents as a single dose form oras separate dosage forms.
 31. A compound of claim 1 or enantiomers,diastereomers, tautomers, and pharmaceutically acceptable salts, whereinsaid compound is selected from: